- Email: [email protected]
Comparison of the clinical performances of the AdvanSure HPV Screening Real-Time PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-Risk HPV DNA Test for Cervical Cancer Screening
Accepted Manuscript Title: Comparison of the Clinical Performances of the AdvanSure HPV Screening Real-Time PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-Risk HPV DNA Test for Cervical Cancer Screening Author: Hae-Sun Chung Chorong Hahm Miae Lee PII: DOI: Reference:
S0166-0934(14)00181-5 http://dx.doi.org/doi:10.1016/j.jviromet.2014.04.021 VIRMET 12514
To appear in:
Journal of Virological Methods
Received date: Revised date: Accepted date:
11-11-2013 24-4-2014 29-4-2014
Please cite this article as: Chung, H.-S., Hahm, C., Lee, M.,Comparison of the Clinical Performances of the AdvanSure HPV Screening Real-Time PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-Risk HPV DNA Test for Cervical Cancer Screening, Journal of Virological Methods (2014), http://dx.doi.org/10.1016/j.jviromet.2014.04.021 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
*Highlights (for review)
Highlights
We evaluated the clinical performance of three HPV DNA assays for detection of cervical intraepithelial neoplasia of grade 2 or worse.
The three HPV DNA commercial assays showed relatively good clinical sensitivity.
The AdvanSure PCR had a lower clinical specificity than the Abbott PCR and the
ip t
HC2. The AdvanSure and the Abbott PCR assays can be useful for detection of HPV
The two real-time PCR assays are useful in HPV testing for cervical cancer
ce pt
ed
M
an
us
screening.
Ac
cr
types 16/18.
Page 1 of 17
Table(s)
1
Table 1. Comparison of clinical sensitivity and specificity for the detection of cervical
2
intraepithelial neoplasia of grade 2 or worse of the HC2, the AdvanSure PCR, and the
3
Abbott PCR assay Cervical
Clinical
Clinical
sensitivity
specificity
(%)
(%)
ip t
intraepithelial neoplasia of
us
worse -
+
22
100
100
-
0
497
(84.4-100)
Abbott
+
21
81
95.5
PCR
-
1
516
(77.1-99.2)
AdvanSure +
21
229
1
368
an
+
83.3
18.0
100
(80.0-86.2)
(11.7-26.0)
(99.3-100)
86.4
20.6
99.8
(83.4-89.1)
(13.2-29.7)
(98.9-100)
95.5
61.6
8.4
99.7
(77.1-99.2)
(57.6-65.6)
(5.3-12.6)
(98.5-100)
ed
ce pt
-
M
HC2
PCR
NPV (%)
cr
PPV (%)
grade 2 or
95% confidence intervals are shown in parentheses. Abbreviations: HC2, Hybrid Capture 2; PPV, positive predictive value; NPV, negative
Ac
predictive value
1
Page 2 of 17
4
Table 2. Concordance between the Abbott PCR and the HC2 for high-risk HPV
5
detection. Abbott PCR
+
+
-
88
34
HC2
0.74 (0.67-0.81) 483
ce pt
ed
M
an
us
Abbreviation: HC2, Hybrid Capture 2
Ac
6
14
92.1%
cr
-
Concordance rate
ip t
kappa (95% CI)
2
Page 3 of 17
7
Table 3. The discordant results and interpretation of the HC2 and the Abbott PCR assays
8
for high-risk HPV detection with respect to the cervical histopathology (cervical
9
intraepithelial neoplasia of grade 2 or worse)
N
Result
Interpretation
Result
1
+
True positive
-
14
-
True negative
+
33
+
False positive
us
cr
False negative False positive True negative
ce pt
ed
M
an
Abbreviation: HC2, Hybrid Capture 2
-
Interpretation
Ac
10
Abbott PCR
ip t
HC2
3
Page 4 of 17
Manuscript
TITLE PAGE
Title: Comparison of the Clinical Performances of the AdvanSure HPV Screening RealTime PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-
cr
ip t
Risk HPV DNA Test for Cervical Cancer Screening
Authors: Hae-Sun Chung, Chorong Hahm, and Miae Lee
us
Running title: Clinical performances of 3 HPV DNA testing assays
an
Department of Laboratory Medicine, Ewha Womans University School of Medicine, 1071
Fax: +82-2-2650-5091 Email: [email protected]
Ac
ce pt
Tel: +82-2-2650-5222
ed
Corresponding author: Miae Lee
M
Anyangcheon-ro, Yangcheon-gu, Seoul, South Korea, 158-710
Page 5 of 17
ASTRACT
2
The clinical performance of three human papillomavirus (HPV) DNA commercial
3
assays for cervical cancer screening was evaluated; the AdvanSure HPV Screening
4
Real-Time PCR (AdvanSure PCR; LG Life Sciences) that was developed recently for
5
the detection of both high-risk and low-risk genotypes, the Abbott RealTime High-Risk
6
HPV Test (Abbott PCR; Abbott Molecular) and the Hybrid Capture High-Risk HPV
7
DNA Test (HC2; Qiagen). The three different HPV DNA tests were compared using
8
cytology samples obtained from 619 women who underwent routine cervical cancer
9
screening. The gold-standard assay was histopathological confirmation of cervical
10
intraepithelial neoplasia of grade 2 or worse. The clinical sensitivities of the AdvanSure
11
PCR, the Abbott PCR and the HC2 for the detection of cervical intraepithelial neoplasia
12
of grade 2 or worse were 95.5%, 95.5% and 100%, respectively, while the clinical
13
specificities were 61.6%, 86.4% and 83.3%, respectively. There were no significant
14
differences in the clinical sensitivities of the Abbott PCR and the AdvanSure PCR
15
compared to the HC2. The clinical specificities of the Abbott PCR and the AdvanSure
16
PCR for the detection of HPV types 16/18 were 97.8% and 98.5%, respectively. For
17
cervical cancer screening, all three tests showed relatively good clinical sensitivities, but
18
the AdvanSure PCR had lower clinical specificity than the Abbott PCR and the HC2.
19
The AdvanSure PCR and the Abbott PCR assays have the advantage of being automated
20
and the ability to distinguish between HPV types 16/18 and other HPV types. The two
21
real-time PCR assays could be useful tools in HPV testing for cervical cancer screening.
22
Keywords: Human papillomavirus (HPV), real-time PCR, clinical sensitivity, clinical
23
specificity, cervical cancer screening
Ac
ce pt
ed
M
an
us
cr
ip t
1
2
Page 6 of 17
24
1. Introduction Persistent human papillomavirus (HPV) infection is a major risk factor in the
26
development of carcinoma of the uterine cervix. DNA-based screening assays for the
27
detection of HPV are significantly more sensitive than Pap cytology for the detection of
28
high-grade cervical intraepithelial neoplasia (cervical intraepithelial neoplasia of grade
29
2 or worse) and are recommended for primary cervical cancer screening and triage of
30
borderline cytological abnormalities (Meijer et al., 2009; Poljak and Kocjan, 2010).
cr
ip t
25
For the last decade, the Hybrid Capture 2 (HC2; Qiagen, Gaithersburg, MD, USA)
32
test has been the most important HPV diagnostic assay, and it is still the used most
33
frequently worldwide. This signal-amplification assay is based on a sandwich capture
34
molecular
35
chemiluminescent detection and can detect 13 high-risk HPV genotypes and 5 low-risk
36
HPV genotypes. The Abbott RealTime High Risk HPV PCR assay (Abbott PCR; Abbott
37
Molecular, Abbott Park, IL, USA) is a real-time PCR assay designed to detect 14 high-
38
risk HPV genotypes and simultaneously distinguish HPV types 16/18 from other HPV
39
types (Poljak and Kocjan, 2010; Abreu et al., 2012). LG Life Sciences (Seoul, Korea)
40
has also developed a real-time PCR assay (AdvanSure HPV Screening real-time PCR;
41
AdvanSure PCR), which detects 41 HPV genotypes and simultaneously differentiates
42
HPV types 16/18 from other HPV types (Hwang and Lee, 2012).
that
provides
a
semiquantitative
result
through
Ac
ce pt
ed
M
hybridization
an
us
31
43
Previously, the analytical performances of the two real-time PCR assays, the
44
AdvanSure PCR and the Abbott PCR, were investigated and compared with the HC2
45
assay (Hwang and Lee, 2012). However, the performance of the AdvanSure PCR has
46
not been fully evaluated yet, especially regarding the clinical performance. In this study,
47
the clinical performance of the three HPV DNA commercial assays for cervical cancer 3
Page 7 of 17
48
screening was evaluated.
49 50
2. Materials and methods
51
2.1. Specimens Cytology samples were obtained from 619 women undergoing routine cervical
53
cancer screening during medical examination at Ewha Womans University Health
54
Promotion Center from October to November 2011. Patients who had been treated for
55
cervical intraepithelial neoplasia prior to this study were excluded. A single cervical
56
specimen was collected from all participants using a cytobrush and was suspended in
57
Cervical Sampler solution (Qiagen, Gaithersburg, MD, USA) according to the
58
manufacturer’s instructions. The pathological results on the basis of the cervical
59
cytology or biopsy results were within the normal limits in 538 patients (86.9%).
60
Atypical squamous cells of undetermined significance were observed in 28 (4.5%),
61
cervical intraepithelial neoplasia grade 1 in 31 (5.0%), cervical intraepithelial neoplasia
62
grade 2 in 8 (1.3%), cervical intraepithelial neoplasia grade 3 in 12 (1.9%) and cervical
63
carcinoma in 2 patients (0.3%).
66
cr
us
an
M
ed
ce pt
65
2.2. Assays for HPV detection
Ac
64
ip t
52
The specimens were analyzed as described previously (Hwang and Lee, 2012). All
67
specimens were tested using the HC2, the Abbott PCR and the AdvanSure PCR assays.
68
HC2 can detect 13 high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58,
69
59 and 68). Specimens with relative light unit/cutoff ratios (RLU/CO) ≥1.0 were
70
considered positive. The Abbott PCR was developed for the detection of DNA from 14
71
high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68). It 4
Page 8 of 17
72
also simultaneously distinguishes HPV types 16/18 from other HPV types. The assays
73
were performed according to the manufacturers’ instructions. The AdvanSure PCR detects 41 HPV genotypes (high-risk: 16, 18, 26, 31, 33, 35,
75
39, 45, 51, 52, 53, 56, 58, 59, 66, 67, 68, 69, 70, 72 and 73; and low-risk: 3, 6, 10, 11,
76
27, 32, 34, 40, 42, 43, 44, 54, 55, 57, 61, 62, 71, 74, 81 and 84) and also differentiates
77
HPV types 16/18 from other HPV types. It utilizes the one-tube nested multiplex PCR
78
technique using nucleic acid intercalating dye and common primer sets for HPV L1
79
regions (de Roda Husman et al., 1995; Karlsen et al., 1996). HPV types 16/18 can be
80
detected separately among several HPV types using HPV types 16/18 specific probes.
81
The Taqman chemistry technique is applied to the internal control and HPV types 16/18.
82
PCR was performed with preliminary denaturation at 95°C for 10 min, followed by 42
83
cycles of denaturation at 95°C for 15 s, annealing at 55°C in the early 10 cycles and at
84
48°C in the later 32 cycles for 45 s, and extension at 74°C for 30 s. The criteria of Ct
85
value for positive are 30 for HPV and 32 for HPV types 16/18. The sensitivity of the
86
AdvanSure PCR is variable depending on HPV types, ranged from 0.005 to 0.369
87
pg/mL. The LODs for HPV types 16 and 18 is 0.037 and 0.072 pg/mL, respectively
88
(according to the instructions provided by the manufacturer).
90 91
cr
us
an
M
ed
ce pt
Ac
89
ip t
74
2.3. Statistical analysis For assessment of the clinical performance of the HC2, the Abbott PCR and the
92
AdvanSure PCR, cases were defined as women with high-grade cervical disease
93
(histopathologically cervical intraepithelial neoplasia grade 2 or worse) and controls as
94
women without high-grade cervical disease (less than cervical intraepithelial neoplasia
95
grade 2). Clinical sensitivity, clinical specificity, negative predictive value (NPV) and 5
Page 9 of 17
positive predictive value (PPV) were calculated using the conventional contingency
97
tables and 95% confidence intervals (95% CI) were computed using exact binomial
98
methods. Clinical sensitivity and specificity of the Abbott PCR and AdvanSure PCR
99
were compared to the HC2 by McNemar’s test. Agreement between assays was assessed
100
by the kappa statistic. Additionally, the results for the detection of HPV types 16/18 of
101
the two real-time PCR assays were also compared.
cr
ip t
96
102
3. Results
us
103
Clinical sensitivities and clinical specificities for the detection of cervical
105
intraepithelial neoplasia of grade 2 or worse of the HC2, the Abbott PCR and the
106
AdvanSure PCR assays are summarized in Table 1. There were no significant
107
differences in the clinical sensitivities of the Abbott PCR and the AdvanSure PCR
108
compared to the HC2 (P=1.0). Clinical specificity of the Abbott PCR was higher than
109
the HC2 (P=0.0087), whereas that of the AdvanSure PCR was significantly lower than
110
the HC2 (P<0.0001).
ce pt
ed
M
an
104
The percent agreement between the HC2 and the Abbott PCR for high-risk HPV
112
detection was 92.1% (kappa 0.74) (Table 2). Among the 48 discordant results, the HC2
113
yielded 33 false positive results and the Abbott PCR yielded 14 false positive and 1
114
false negative results, with respect to the cervical histopathology (Table 3). The
115
agreements between the AdvanSure PCR and the HC2 and between the AdvanSure PCR
116
and the Abbott PCR were not evaluated, because the AdvanSure PCR detected both
117
high-risk and low-risk HPV.
Ac
111
118
The two real-time PCR assays could separately detect oncogenic, high-risk HPV
119
types 16/18. Among 619 patients, HPV types 16/18 were detected in 28 and 26 patients 6
Page 10 of 17
120
by the AdvanSure PCR and the Abbott PCR, respectively. They showed excellent
121
agreement (99.4% agreement, kappa 0.92). The clinical specificities of the detection of
122
HPV types 16/18 for the detection of cervical intraepithelial neoplasia of grade 2 or
123
worse by the AdvanSure PCR and the Abbott PCR were 98.5% and 97.8%, respectively.
4. Discussion
cr
125
ip t
124
It has been recommended that new HPV assays should have similar clinical
127
characteristics to the HC2 in the process of clinical validation of high-risk HPV tests
128
before they can be used in cervical cancer screening (Meijer et al., 2009). Previous
129
study which had investigated the analytical sensitivities and specificities of the
130
AdvanSure PCR and the Abbott PCR for HPV detection and compared them with the
131
HC2 revealed that the AdvanSure PCR and the Abbott PCR were less sensitive but more
132
specific than the HC2 for the detection of HPV, and the overall performance of the two
133
real-time PCR assays was comparable to the HC2 (Hwang and Lee, 2012).
ed
M
an
us
126
For the detection of cervical intraepithelial neoplasia of grade 2 or worse, all three
135
tests showed relatively good clinical sensitivities (95.5-100%), but the AdvanSure PCR
136
had a relatively lower clinical specificity (61.6%) compared with the HC2 (83.3%) and
137
the Abbott PCR (86.4%). This is probably because the AdvanSure PCR can detect both
138
high-risk and low-risk HPV. It is now well-established that virtually all cervical cancer
139
and its immediate pre-cancerous lesions arise from persisting cervical infections by
140
high-risk HPV including HPV types 16/18 (Munoz et al., 2003; Smith et al., 2007;
141
Bosch et al., 2008; de Freitas et al., 2012). On the contrary, low-risk types are rarely
142
associated with cancer. Therefore, most of the positive results of the AdvanSure PCR
143
due to low-risk HPV were likely to be false positive results. These results are similar to
Ac
ce pt
134
7
Page 11 of 17
data shown in a previous study (Choi et al., 2013); the AdvanSure PCR showed high
145
numbers of positive results in the normal cytological group, the atypical squamous cells
146
of undetermined significance group and the low-grade squamous intraepithelial lesion
147
group (88.5%, 70.6% and 93.5%, respectively), indicating a low clinical specificity of
148
the assay because all of them were false positive results for the detection of cervical
149
intraepithelial neoplasia of grade 2 or worse. Most of the false positive results were due
150
to HPV types other than 16/18. On the other hand, the HC2 and the Abbott PCR showed
151
relatively low positive rates; the positive rates of normal, atypical squamous cells of
152
undetermined significance and low-grade squamous intraepithelial lesion group were
153
10.3%, 25.5% and 65.2%, respectively by the HC2, and those were 14.1%, 27.5% and
154
50.0%, respectively by the Abbott PCR. Further studies are necessary to determine the
155
clinical usefulness of the detection of low-risk HPV.
M
an
us
cr
ip t
144
Literature data on clinical validation of the Abbott RCR showed high clinical
157
sensitivity for the detection of cervical intraepithelial neoplasia of grade 2 or worse
158
(over 90% in the most studies) and it was comparable with the HC2 (Huang et al., 2009;
159
Tang et al., 2009; Cuzick et al., 2010; Halfon et al., 2010; Carozzi et al., 2011; Poljak et
160
al., 2011). But the clinical specificity was variable; about 90% in some studies (Carozzi
161
et al., 2011; Poljak et al., 2011) and less than 50% in other studies (Huang et al., 2009;
162
Cuzick et al., 2010; Halfon et al., 2010). This variance may be due to difference in study
163
populations. Disease prevalence of study population is an important consideration for
164
interpreting the predictive value of a test. The clinical sensitivity and specificity of the
165
Abbott PCR in this study were similar to the previous results (Poljak et al., 2009;
166
Halfon et al., 2010; Carozzi et al., 2011; Poljak et al., 2011).
167
Ac
ce pt
ed
156
The AdvanSure PCR and the Abbott PCR were evaluated comparing to the HC2. 8
Page 12 of 17
However, in this study, the sample size was smaller than recommended sample size for
169
the validation of the clinical sensitivity and specificity by the noninferiority test (Meijer
170
et al., 2009). Therefore, interpretation of the results had limitation. Even the Abbott
171
PCR and the AdvanSure PCR showed relatively high clinical sensitivities (both 95.5%),
172
but noninferiority to the HC2 was not demonstrated in this study (data now shown). It
173
might be due to the small sample size. But the clinical sensitivities of the AdvanSure
174
PCR and the Abbott PCR were over 95% (even though the confidence intervals were
175
wide), and there were no significant differences in the clinical sensitivities of the Abbott
176
PCR and the AdvanSure PCR compared to the HC2. While the AdvanSure PCR had a
177
relatively lower clinical specificity compared with the HC2, the Abbott PCR showed a
178
good agreement with the HC2 and a higher clinical specificity for the detection of
179
cervical intraepithelial neoplasia of grade 2 or worse compared with the HC2.
M
an
us
cr
ip t
168
The two real-time assays are expected to be useful for the detection of HPV types
181
16/18 in cervical cancer screening. HPV types 16/18 are the most important genotypes,
182
and account for about 70% of all invasive cervical cancers (Smith et al., 2007).
183
Therefore, the clinical specificity of the detection of HPV types 16/18 is highly likely to
184
be excellent, as shown in this study. And the detection of HPV types 16/18 is significant
185
in itself, regardless of the current status, because patients with these HPV genotypes
186
have a high chance of developing high-grade cervical disease. However, high-risk HPV
187
infection is not sufficient by itself to cause cervical cancer and a large number of the
188
women infected with high-risk HPV will not develop cervical disease (de Freitas et al.,
189
2012), so careful thought and consideration are needed for the cases in the detection of
190
high-risk HPV with normal cytology. If HPV types 16 or 18 is detected in women who
191
are cytology-negative, referral to a gynecologic oncologist and a colposcopic
Ac
ce pt
ed
180
9
Page 13 of 17
192
examination are recommended (Lee et al., 2013; Saslow et al., 2012). In summary, for cervical cancer screening, all three tests showed relatively good
194
clinical sensitivities, but the AdvanSure PCR had a lower clinical specificity than the
195
Abbott PCR and the HC2. The Abbott PCR and the AdvanSure PCR have the advantage
196
of being automated and the ability to distinguish between HPV types 16/18 and other
197
HPV types. They could be useful tools in HPV testing for cervical cancer screening.
cr
ip t
193
198
Acknowledgements
us
199
Abbott Korea (Seoul, Korea) and LG Life Sciences (Seoul, Korea) provided with
201
the equipments and reagents for this study. We would like to thank to Kyoung Ae Kong,
202
Clinical Trial Center, Ewha Womans University Medical Center, for statistical support
203
for this study.
Ac
ce pt
ed
M
an
200
10
Page 14 of 17
204
References
205
207
Abreu, A.L., Souza, R.P., Gimenes, F., Consolaro, M.E., 2012. A review of methods for detect human Papillomavirus infection. Virol J 9, 262.
ip t
206
Bosch, F.X., Burchell, A.N., Schiffman, M., Giuliano, A.R., de Sanjose, S., Bruni, L.,
209
Tortolero-Luna, G., Kjaer, S.K., Munoz, N., 2008. Epidemiology and natural
210
history of human papillomavirus infections and type-specific implications in
211
cervical neoplasia. Vaccine 26 Suppl 10, K1-16.
us
cr
208
Carozzi, F.M., Burroni, E., Bisanzi, S., Puliti, D., Confortini, M., Giorgi Rossi, P., Sani,
213
C., Scalisi, A., Chini, F., 2011. Comparison of clinical performance of Abbott
214
RealTime High Risk HPV test with that of hybrid capture 2 assay in a screening
215
setting. J Clin Microbiol 49, 1446-1451.
M
an
212
Choi, J., Park, Y., Lee, E.H., Kim, S., Kim, J.H., Kim, H.S., 2013. Detection and
217
genotyping of human papillomavirus by five assays according to cytologic results.
218
J Virol Methods 187, 79-84.
ed
216
Cuzick, J., Ambroisine, L., Cadman, L., Austin, J., Ho, L., Terry, G., Liddle, S., Dina, R.,
220
McCarthy, J., Buckley, H., et al., 2010. Performance of the Abbott RealTime high-
221
risk HPV test in women with abnormal cervical cytology smears. J Med Virol 82,
222
1186-1191.
Ac
ce pt
219
223
de Freitas, A.C., Gurgel, A.P., Chagas, B.S., Coimbra, E.C., do Amaral, C.M., 2012.
224
Susceptibility to cervical cancer: an overview. Gynecol Oncol 126, 304-311.
225
de Roda Husman, A.M., Walboomers, J.M., van den Brule, A.J., Meijer, C.J., Snijders,
226
P.J., 1995. The use of general primers GP5 and GP6 elongated at their 3' ends with
227
adjacent highly conserved sequences improves human papillomavirus detection
228
by PCR. J Gen Virol 76 ( Pt 4), 1057-1062.
229
Halfon, P., Benmoura, D., Agostini, A., Khiri, H., Penaranda, G., Martineau, A., Blanc, 11
Page 15 of 17
230
B., 2010. Evaluation of the clinical performance of the Abbott RealTime High-
231
Risk HPV for carcinogenic HPV detection. J Clin Virol 48, 246-250. Huang, S., Erickson, B., Tang, N., Mak, W.B., Salituro, J., Robinson, J., Abravaya, K.,
233
2009. Clinical performance of Abbott RealTime High Risk HPV test for detection
234
of high-grade cervical intraepithelial neoplasia in women with abnormal cytology.
235
J Clin Virol 45 Suppl 1, S19-23.
ip t
232
Hwang, Y., Lee, M., 2012. Comparison of the AdvanSure human papillomavirus
237
screening real-time PCR, the Abbott RealTime High Risk human papillomavirus
238
test, and the Hybrid Capture human papillomavirus DNA test for the detection of
239
human papillomavirus. Ann Lab Med 32, 201-205.
an
us
cr
236
Karlsen, F., Kalantari, M., Jenkins, A., Pettersen, E., Kristensen, G., Holm, R.,
241
Johansson, B., Hagmar, B., 1996. Use of multiple PCR primer sets for optimal
242
detection of human papillomavirus. J Clin Microbiol 34, 2095-2100.
M
240
Lee, J.K., Hong, J.H., Kang, S., Kim, D.Y., Kim, B.G., Kim, S.H., Kim, Y.M., Kim,
244
J.W., Kim, J.H., Kim, T.J., et al., 2013. Practice guidelines for the early detection
245
of cervical cancer in Korea: Korean Society of Gynecologic Oncology and the
246
Korean Society for Cytopathology 2012 edition. J Gynecol Oncol 24, 186-203.
ce pt
ed
243
Meijer, C.J., Berkhof, J., Castle, P.E., Hesselink, A.T., Franco, E.L., Ronco, G., Arbyn,
248
M., Bosch, F.X., Cuzick, J., Dillner, J., et al., 2009. Guidelines for human
249
papillomavirus DNA test requirements for primary cervical cancer screening in
250
Ac
247
women 30 years and older. Int J Cancer 124, 516-520.
251
Munoz, N., Bosch, F.X., de Sanjose, S., Herrero, R., Castellsague, X., Shah, K.V.,
252
Snijders, P.J., Meijer, C.J., 2003. Epidemiologic classification of human
253
papillomavirus types associated with cervical cancer. N Engl J Med 348, 518-527.
254
Poljak, M., Kovanda, A., Kocjan, B.J., Seme, K., Jancar, N., Vrtacnik-Bokal, E., 2009.
255
The Abbott RealTime High Risk HPV test: comparative evaluation of analytical 12
Page 16 of 17
256
specificity and clinical sensitivity for cervical carcinoma and CIN 3 lesions with
257
the Hybrid Capture 2 HPV DNA test. Acta Dermatovenerol Alp Panonica Adriat
258
18, 94-103. Poljak, M., Kocjan, B.J., 2010. Commercially available assays for multiplex detection
260
of alpha human papillomaviruses. Expert Rev Anti Infect Ther 8, 1139-1162.
261
Poljak, M., Ostrbenk, A., Seme, K., Ucakar, V., Hillemanns, P., Bokal, E.V., Jancar, N.,
262
Klavs, I., 2011. Comparison of clinical and analytical performance of the Abbott
263
Realtime High Risk HPV test to the performance of hybrid capture 2 in
264
population-based cervical cancer screening. J Clin Microbiol 49, 1721-1729.
265
Saslow, D., Solomon, D., Lawson, H.W., Killackey, M., Kulasingam, S.L., Cain, J.M.,
266
Garcia, F.A., Moriarty, A.T., Waxman, A.G., Wilbur, D.C., et al., 2012. American
267
Cancer Society, American Society for Colposcopy and Cervical Pathology, and
268
American Society for Clinical Pathology screening guidelines for the prevention
269
and early detection of cervical cancer. J Low Genit Tract Dis 16, 175-204.
ed
M
an
us
cr
ip t
259
Smith, J.S., Lindsay, L., Hoots, B., Keys, J., Franceschi, S., Winer, R., Clifford, G.M.,
271
2007. Human papillomavirus type distribution in invasive cervical cancer and
272
high-grade cervical lesions: a meta-analysis update. Int J Cancer 121, 621-632.
ce pt
270
Tang, N., Huang, S., Erickson, B., Mak, W.B., Salituro, J., Robinson, J., Abravaya, K.,
274
2009. High-risk HPV detection and concurrent HPV 16 and 18 typing with Abbott
275
RealTime High Risk HPV test. J Clin Virol 45 Suppl 1, S25-28.
Ac
273
13
Page 17 of 17
S0166-0934(14)00181-5 http://dx.doi.org/doi:10.1016/j.jviromet.2014.04.021 VIRMET 12514
To appear in:
Journal of Virological Methods
Received date: Revised date: Accepted date:
11-11-2013 24-4-2014 29-4-2014
Please cite this article as: Chung, H.-S., Hahm, C., Lee, M.,Comparison of the Clinical Performances of the AdvanSure HPV Screening Real-Time PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-Risk HPV DNA Test for Cervical Cancer Screening, Journal of Virological Methods (2014), http://dx.doi.org/10.1016/j.jviromet.2014.04.021 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
*Highlights (for review)
Highlights
We evaluated the clinical performance of three HPV DNA assays for detection of cervical intraepithelial neoplasia of grade 2 or worse.
The three HPV DNA commercial assays showed relatively good clinical sensitivity.
The AdvanSure PCR had a lower clinical specificity than the Abbott PCR and the
ip t
HC2. The AdvanSure and the Abbott PCR assays can be useful for detection of HPV
The two real-time PCR assays are useful in HPV testing for cervical cancer
ce pt
ed
M
an
us
screening.
Ac
cr
types 16/18.
Page 1 of 17
Table(s)
1
Table 1. Comparison of clinical sensitivity and specificity for the detection of cervical
2
intraepithelial neoplasia of grade 2 or worse of the HC2, the AdvanSure PCR, and the
3
Abbott PCR assay Cervical
Clinical
Clinical
sensitivity
specificity
(%)
(%)
ip t
intraepithelial neoplasia of
us
worse -
+
22
100
100
-
0
497
(84.4-100)
Abbott
+
21
81
95.5
PCR
-
1
516
(77.1-99.2)
AdvanSure +
21
229
1
368
an
+
83.3
18.0
100
(80.0-86.2)
(11.7-26.0)
(99.3-100)
86.4
20.6
99.8
(83.4-89.1)
(13.2-29.7)
(98.9-100)
95.5
61.6
8.4
99.7
(77.1-99.2)
(57.6-65.6)
(5.3-12.6)
(98.5-100)
ed
ce pt
-
M
HC2
PCR
NPV (%)
cr
PPV (%)
grade 2 or
95% confidence intervals are shown in parentheses. Abbreviations: HC2, Hybrid Capture 2; PPV, positive predictive value; NPV, negative
Ac
predictive value
1
Page 2 of 17
4
Table 2. Concordance between the Abbott PCR and the HC2 for high-risk HPV
5
detection. Abbott PCR
+
+
-
88
34
HC2
0.74 (0.67-0.81) 483
ce pt
ed
M
an
us
Abbreviation: HC2, Hybrid Capture 2
Ac
6
14
92.1%
cr
-
Concordance rate
ip t
kappa (95% CI)
2
Page 3 of 17
7
Table 3. The discordant results and interpretation of the HC2 and the Abbott PCR assays
8
for high-risk HPV detection with respect to the cervical histopathology (cervical
9
intraepithelial neoplasia of grade 2 or worse)
N
Result
Interpretation
Result
1
+
True positive
-
14
-
True negative
+
33
+
False positive
us
cr
False negative False positive True negative
ce pt
ed
M
an
Abbreviation: HC2, Hybrid Capture 2
-
Interpretation
Ac
10
Abbott PCR
ip t
HC2
3
Page 4 of 17
Manuscript
TITLE PAGE
Title: Comparison of the Clinical Performances of the AdvanSure HPV Screening RealTime PCR, the Abbott Real-Time High-Risk HPV Test, and the Hybrid Capture High-
cr
ip t
Risk HPV DNA Test for Cervical Cancer Screening
Authors: Hae-Sun Chung, Chorong Hahm, and Miae Lee
us
Running title: Clinical performances of 3 HPV DNA testing assays
an
Department of Laboratory Medicine, Ewha Womans University School of Medicine, 1071
Fax: +82-2-2650-5091 Email: [email protected]
Ac
ce pt
Tel: +82-2-2650-5222
ed
Corresponding author: Miae Lee
M
Anyangcheon-ro, Yangcheon-gu, Seoul, South Korea, 158-710
Page 5 of 17
ASTRACT
2
The clinical performance of three human papillomavirus (HPV) DNA commercial
3
assays for cervical cancer screening was evaluated; the AdvanSure HPV Screening
4
Real-Time PCR (AdvanSure PCR; LG Life Sciences) that was developed recently for
5
the detection of both high-risk and low-risk genotypes, the Abbott RealTime High-Risk
6
HPV Test (Abbott PCR; Abbott Molecular) and the Hybrid Capture High-Risk HPV
7
DNA Test (HC2; Qiagen). The three different HPV DNA tests were compared using
8
cytology samples obtained from 619 women who underwent routine cervical cancer
9
screening. The gold-standard assay was histopathological confirmation of cervical
10
intraepithelial neoplasia of grade 2 or worse. The clinical sensitivities of the AdvanSure
11
PCR, the Abbott PCR and the HC2 for the detection of cervical intraepithelial neoplasia
12
of grade 2 or worse were 95.5%, 95.5% and 100%, respectively, while the clinical
13
specificities were 61.6%, 86.4% and 83.3%, respectively. There were no significant
14
differences in the clinical sensitivities of the Abbott PCR and the AdvanSure PCR
15
compared to the HC2. The clinical specificities of the Abbott PCR and the AdvanSure
16
PCR for the detection of HPV types 16/18 were 97.8% and 98.5%, respectively. For
17
cervical cancer screening, all three tests showed relatively good clinical sensitivities, but
18
the AdvanSure PCR had lower clinical specificity than the Abbott PCR and the HC2.
19
The AdvanSure PCR and the Abbott PCR assays have the advantage of being automated
20
and the ability to distinguish between HPV types 16/18 and other HPV types. The two
21
real-time PCR assays could be useful tools in HPV testing for cervical cancer screening.
22
Keywords: Human papillomavirus (HPV), real-time PCR, clinical sensitivity, clinical
23
specificity, cervical cancer screening
Ac
ce pt
ed
M
an
us
cr
ip t
1
2
Page 6 of 17
24
1. Introduction Persistent human papillomavirus (HPV) infection is a major risk factor in the
26
development of carcinoma of the uterine cervix. DNA-based screening assays for the
27
detection of HPV are significantly more sensitive than Pap cytology for the detection of
28
high-grade cervical intraepithelial neoplasia (cervical intraepithelial neoplasia of grade
29
2 or worse) and are recommended for primary cervical cancer screening and triage of
30
borderline cytological abnormalities (Meijer et al., 2009; Poljak and Kocjan, 2010).
cr
ip t
25
For the last decade, the Hybrid Capture 2 (HC2; Qiagen, Gaithersburg, MD, USA)
32
test has been the most important HPV diagnostic assay, and it is still the used most
33
frequently worldwide. This signal-amplification assay is based on a sandwich capture
34
molecular
35
chemiluminescent detection and can detect 13 high-risk HPV genotypes and 5 low-risk
36
HPV genotypes. The Abbott RealTime High Risk HPV PCR assay (Abbott PCR; Abbott
37
Molecular, Abbott Park, IL, USA) is a real-time PCR assay designed to detect 14 high-
38
risk HPV genotypes and simultaneously distinguish HPV types 16/18 from other HPV
39
types (Poljak and Kocjan, 2010; Abreu et al., 2012). LG Life Sciences (Seoul, Korea)
40
has also developed a real-time PCR assay (AdvanSure HPV Screening real-time PCR;
41
AdvanSure PCR), which detects 41 HPV genotypes and simultaneously differentiates
42
HPV types 16/18 from other HPV types (Hwang and Lee, 2012).
that
provides
a
semiquantitative
result
through
Ac
ce pt
ed
M
hybridization
an
us
31
43
Previously, the analytical performances of the two real-time PCR assays, the
44
AdvanSure PCR and the Abbott PCR, were investigated and compared with the HC2
45
assay (Hwang and Lee, 2012). However, the performance of the AdvanSure PCR has
46
not been fully evaluated yet, especially regarding the clinical performance. In this study,
47
the clinical performance of the three HPV DNA commercial assays for cervical cancer 3
Page 7 of 17
48
screening was evaluated.
49 50
2. Materials and methods
51
2.1. Specimens Cytology samples were obtained from 619 women undergoing routine cervical
53
cancer screening during medical examination at Ewha Womans University Health
54
Promotion Center from October to November 2011. Patients who had been treated for
55
cervical intraepithelial neoplasia prior to this study were excluded. A single cervical
56
specimen was collected from all participants using a cytobrush and was suspended in
57
Cervical Sampler solution (Qiagen, Gaithersburg, MD, USA) according to the
58
manufacturer’s instructions. The pathological results on the basis of the cervical
59
cytology or biopsy results were within the normal limits in 538 patients (86.9%).
60
Atypical squamous cells of undetermined significance were observed in 28 (4.5%),
61
cervical intraepithelial neoplasia grade 1 in 31 (5.0%), cervical intraepithelial neoplasia
62
grade 2 in 8 (1.3%), cervical intraepithelial neoplasia grade 3 in 12 (1.9%) and cervical
63
carcinoma in 2 patients (0.3%).
66
cr
us
an
M
ed
ce pt
65
2.2. Assays for HPV detection
Ac
64
ip t
52
The specimens were analyzed as described previously (Hwang and Lee, 2012). All
67
specimens were tested using the HC2, the Abbott PCR and the AdvanSure PCR assays.
68
HC2 can detect 13 high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58,
69
59 and 68). Specimens with relative light unit/cutoff ratios (RLU/CO) ≥1.0 were
70
considered positive. The Abbott PCR was developed for the detection of DNA from 14
71
high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68). It 4
Page 8 of 17
72
also simultaneously distinguishes HPV types 16/18 from other HPV types. The assays
73
were performed according to the manufacturers’ instructions. The AdvanSure PCR detects 41 HPV genotypes (high-risk: 16, 18, 26, 31, 33, 35,
75
39, 45, 51, 52, 53, 56, 58, 59, 66, 67, 68, 69, 70, 72 and 73; and low-risk: 3, 6, 10, 11,
76
27, 32, 34, 40, 42, 43, 44, 54, 55, 57, 61, 62, 71, 74, 81 and 84) and also differentiates
77
HPV types 16/18 from other HPV types. It utilizes the one-tube nested multiplex PCR
78
technique using nucleic acid intercalating dye and common primer sets for HPV L1
79
regions (de Roda Husman et al., 1995; Karlsen et al., 1996). HPV types 16/18 can be
80
detected separately among several HPV types using HPV types 16/18 specific probes.
81
The Taqman chemistry technique is applied to the internal control and HPV types 16/18.
82
PCR was performed with preliminary denaturation at 95°C for 10 min, followed by 42
83
cycles of denaturation at 95°C for 15 s, annealing at 55°C in the early 10 cycles and at
84
48°C in the later 32 cycles for 45 s, and extension at 74°C for 30 s. The criteria of Ct
85
value for positive are 30 for HPV and 32 for HPV types 16/18. The sensitivity of the
86
AdvanSure PCR is variable depending on HPV types, ranged from 0.005 to 0.369
87
pg/mL. The LODs for HPV types 16 and 18 is 0.037 and 0.072 pg/mL, respectively
88
(according to the instructions provided by the manufacturer).
90 91
cr
us
an
M
ed
ce pt
Ac
89
ip t
74
2.3. Statistical analysis For assessment of the clinical performance of the HC2, the Abbott PCR and the
92
AdvanSure PCR, cases were defined as women with high-grade cervical disease
93
(histopathologically cervical intraepithelial neoplasia grade 2 or worse) and controls as
94
women without high-grade cervical disease (less than cervical intraepithelial neoplasia
95
grade 2). Clinical sensitivity, clinical specificity, negative predictive value (NPV) and 5
Page 9 of 17
positive predictive value (PPV) were calculated using the conventional contingency
97
tables and 95% confidence intervals (95% CI) were computed using exact binomial
98
methods. Clinical sensitivity and specificity of the Abbott PCR and AdvanSure PCR
99
were compared to the HC2 by McNemar’s test. Agreement between assays was assessed
100
by the kappa statistic. Additionally, the results for the detection of HPV types 16/18 of
101
the two real-time PCR assays were also compared.
cr
ip t
96
102
3. Results
us
103
Clinical sensitivities and clinical specificities for the detection of cervical
105
intraepithelial neoplasia of grade 2 or worse of the HC2, the Abbott PCR and the
106
AdvanSure PCR assays are summarized in Table 1. There were no significant
107
differences in the clinical sensitivities of the Abbott PCR and the AdvanSure PCR
108
compared to the HC2 (P=1.0). Clinical specificity of the Abbott PCR was higher than
109
the HC2 (P=0.0087), whereas that of the AdvanSure PCR was significantly lower than
110
the HC2 (P<0.0001).
ce pt
ed
M
an
104
The percent agreement between the HC2 and the Abbott PCR for high-risk HPV
112
detection was 92.1% (kappa 0.74) (Table 2). Among the 48 discordant results, the HC2
113
yielded 33 false positive results and the Abbott PCR yielded 14 false positive and 1
114
false negative results, with respect to the cervical histopathology (Table 3). The
115
agreements between the AdvanSure PCR and the HC2 and between the AdvanSure PCR
116
and the Abbott PCR were not evaluated, because the AdvanSure PCR detected both
117
high-risk and low-risk HPV.
Ac
111
118
The two real-time PCR assays could separately detect oncogenic, high-risk HPV
119
types 16/18. Among 619 patients, HPV types 16/18 were detected in 28 and 26 patients 6
Page 10 of 17
120
by the AdvanSure PCR and the Abbott PCR, respectively. They showed excellent
121
agreement (99.4% agreement, kappa 0.92). The clinical specificities of the detection of
122
HPV types 16/18 for the detection of cervical intraepithelial neoplasia of grade 2 or
123
worse by the AdvanSure PCR and the Abbott PCR were 98.5% and 97.8%, respectively.
4. Discussion
cr
125
ip t
124
It has been recommended that new HPV assays should have similar clinical
127
characteristics to the HC2 in the process of clinical validation of high-risk HPV tests
128
before they can be used in cervical cancer screening (Meijer et al., 2009). Previous
129
study which had investigated the analytical sensitivities and specificities of the
130
AdvanSure PCR and the Abbott PCR for HPV detection and compared them with the
131
HC2 revealed that the AdvanSure PCR and the Abbott PCR were less sensitive but more
132
specific than the HC2 for the detection of HPV, and the overall performance of the two
133
real-time PCR assays was comparable to the HC2 (Hwang and Lee, 2012).
ed
M
an
us
126
For the detection of cervical intraepithelial neoplasia of grade 2 or worse, all three
135
tests showed relatively good clinical sensitivities (95.5-100%), but the AdvanSure PCR
136
had a relatively lower clinical specificity (61.6%) compared with the HC2 (83.3%) and
137
the Abbott PCR (86.4%). This is probably because the AdvanSure PCR can detect both
138
high-risk and low-risk HPV. It is now well-established that virtually all cervical cancer
139
and its immediate pre-cancerous lesions arise from persisting cervical infections by
140
high-risk HPV including HPV types 16/18 (Munoz et al., 2003; Smith et al., 2007;
141
Bosch et al., 2008; de Freitas et al., 2012). On the contrary, low-risk types are rarely
142
associated with cancer. Therefore, most of the positive results of the AdvanSure PCR
143
due to low-risk HPV were likely to be false positive results. These results are similar to
Ac
ce pt
134
7
Page 11 of 17
data shown in a previous study (Choi et al., 2013); the AdvanSure PCR showed high
145
numbers of positive results in the normal cytological group, the atypical squamous cells
146
of undetermined significance group and the low-grade squamous intraepithelial lesion
147
group (88.5%, 70.6% and 93.5%, respectively), indicating a low clinical specificity of
148
the assay because all of them were false positive results for the detection of cervical
149
intraepithelial neoplasia of grade 2 or worse. Most of the false positive results were due
150
to HPV types other than 16/18. On the other hand, the HC2 and the Abbott PCR showed
151
relatively low positive rates; the positive rates of normal, atypical squamous cells of
152
undetermined significance and low-grade squamous intraepithelial lesion group were
153
10.3%, 25.5% and 65.2%, respectively by the HC2, and those were 14.1%, 27.5% and
154
50.0%, respectively by the Abbott PCR. Further studies are necessary to determine the
155
clinical usefulness of the detection of low-risk HPV.
M
an
us
cr
ip t
144
Literature data on clinical validation of the Abbott RCR showed high clinical
157
sensitivity for the detection of cervical intraepithelial neoplasia of grade 2 or worse
158
(over 90% in the most studies) and it was comparable with the HC2 (Huang et al., 2009;
159
Tang et al., 2009; Cuzick et al., 2010; Halfon et al., 2010; Carozzi et al., 2011; Poljak et
160
al., 2011). But the clinical specificity was variable; about 90% in some studies (Carozzi
161
et al., 2011; Poljak et al., 2011) and less than 50% in other studies (Huang et al., 2009;
162
Cuzick et al., 2010; Halfon et al., 2010). This variance may be due to difference in study
163
populations. Disease prevalence of study population is an important consideration for
164
interpreting the predictive value of a test. The clinical sensitivity and specificity of the
165
Abbott PCR in this study were similar to the previous results (Poljak et al., 2009;
166
Halfon et al., 2010; Carozzi et al., 2011; Poljak et al., 2011).
167
Ac
ce pt
ed
156
The AdvanSure PCR and the Abbott PCR were evaluated comparing to the HC2. 8
Page 12 of 17
However, in this study, the sample size was smaller than recommended sample size for
169
the validation of the clinical sensitivity and specificity by the noninferiority test (Meijer
170
et al., 2009). Therefore, interpretation of the results had limitation. Even the Abbott
171
PCR and the AdvanSure PCR showed relatively high clinical sensitivities (both 95.5%),
172
but noninferiority to the HC2 was not demonstrated in this study (data now shown). It
173
might be due to the small sample size. But the clinical sensitivities of the AdvanSure
174
PCR and the Abbott PCR were over 95% (even though the confidence intervals were
175
wide), and there were no significant differences in the clinical sensitivities of the Abbott
176
PCR and the AdvanSure PCR compared to the HC2. While the AdvanSure PCR had a
177
relatively lower clinical specificity compared with the HC2, the Abbott PCR showed a
178
good agreement with the HC2 and a higher clinical specificity for the detection of
179
cervical intraepithelial neoplasia of grade 2 or worse compared with the HC2.
M
an
us
cr
ip t
168
The two real-time assays are expected to be useful for the detection of HPV types
181
16/18 in cervical cancer screening. HPV types 16/18 are the most important genotypes,
182
and account for about 70% of all invasive cervical cancers (Smith et al., 2007).
183
Therefore, the clinical specificity of the detection of HPV types 16/18 is highly likely to
184
be excellent, as shown in this study. And the detection of HPV types 16/18 is significant
185
in itself, regardless of the current status, because patients with these HPV genotypes
186
have a high chance of developing high-grade cervical disease. However, high-risk HPV
187
infection is not sufficient by itself to cause cervical cancer and a large number of the
188
women infected with high-risk HPV will not develop cervical disease (de Freitas et al.,
189
2012), so careful thought and consideration are needed for the cases in the detection of
190
high-risk HPV with normal cytology. If HPV types 16 or 18 is detected in women who
191
are cytology-negative, referral to a gynecologic oncologist and a colposcopic
Ac
ce pt
ed
180
9
Page 13 of 17
192
examination are recommended (Lee et al., 2013; Saslow et al., 2012). In summary, for cervical cancer screening, all three tests showed relatively good
194
clinical sensitivities, but the AdvanSure PCR had a lower clinical specificity than the
195
Abbott PCR and the HC2. The Abbott PCR and the AdvanSure PCR have the advantage
196
of being automated and the ability to distinguish between HPV types 16/18 and other
197
HPV types. They could be useful tools in HPV testing for cervical cancer screening.
cr
ip t
193
198
Acknowledgements
us
199
Abbott Korea (Seoul, Korea) and LG Life Sciences (Seoul, Korea) provided with
201
the equipments and reagents for this study. We would like to thank to Kyoung Ae Kong,
202
Clinical Trial Center, Ewha Womans University Medical Center, for statistical support
203
for this study.
Ac
ce pt
ed
M
an
200
10
Page 14 of 17
204
References
205
207
Abreu, A.L., Souza, R.P., Gimenes, F., Consolaro, M.E., 2012. A review of methods for detect human Papillomavirus infection. Virol J 9, 262.
ip t
206
Bosch, F.X., Burchell, A.N., Schiffman, M., Giuliano, A.R., de Sanjose, S., Bruni, L.,
209
Tortolero-Luna, G., Kjaer, S.K., Munoz, N., 2008. Epidemiology and natural
210
history of human papillomavirus infections and type-specific implications in
211
cervical neoplasia. Vaccine 26 Suppl 10, K1-16.
us
cr
208
Carozzi, F.M., Burroni, E., Bisanzi, S., Puliti, D., Confortini, M., Giorgi Rossi, P., Sani,
213
C., Scalisi, A., Chini, F., 2011. Comparison of clinical performance of Abbott
214
RealTime High Risk HPV test with that of hybrid capture 2 assay in a screening
215
setting. J Clin Microbiol 49, 1446-1451.
M
an
212
Choi, J., Park, Y., Lee, E.H., Kim, S., Kim, J.H., Kim, H.S., 2013. Detection and
217
genotyping of human papillomavirus by five assays according to cytologic results.
218
J Virol Methods 187, 79-84.
ed
216
Cuzick, J., Ambroisine, L., Cadman, L., Austin, J., Ho, L., Terry, G., Liddle, S., Dina, R.,
220
McCarthy, J., Buckley, H., et al., 2010. Performance of the Abbott RealTime high-
221
risk HPV test in women with abnormal cervical cytology smears. J Med Virol 82,
222
1186-1191.
Ac
ce pt
219
223
de Freitas, A.C., Gurgel, A.P., Chagas, B.S., Coimbra, E.C., do Amaral, C.M., 2012.
224
Susceptibility to cervical cancer: an overview. Gynecol Oncol 126, 304-311.
225
de Roda Husman, A.M., Walboomers, J.M., van den Brule, A.J., Meijer, C.J., Snijders,
226
P.J., 1995. The use of general primers GP5 and GP6 elongated at their 3' ends with
227
adjacent highly conserved sequences improves human papillomavirus detection
228
by PCR. J Gen Virol 76 ( Pt 4), 1057-1062.
229
Halfon, P., Benmoura, D., Agostini, A., Khiri, H., Penaranda, G., Martineau, A., Blanc, 11
Page 15 of 17
230
B., 2010. Evaluation of the clinical performance of the Abbott RealTime High-
231
Risk HPV for carcinogenic HPV detection. J Clin Virol 48, 246-250. Huang, S., Erickson, B., Tang, N., Mak, W.B., Salituro, J., Robinson, J., Abravaya, K.,
233
2009. Clinical performance of Abbott RealTime High Risk HPV test for detection
234
of high-grade cervical intraepithelial neoplasia in women with abnormal cytology.
235
J Clin Virol 45 Suppl 1, S19-23.
ip t
232
Hwang, Y., Lee, M., 2012. Comparison of the AdvanSure human papillomavirus
237
screening real-time PCR, the Abbott RealTime High Risk human papillomavirus
238
test, and the Hybrid Capture human papillomavirus DNA test for the detection of
239
human papillomavirus. Ann Lab Med 32, 201-205.
an
us
cr
236
Karlsen, F., Kalantari, M., Jenkins, A., Pettersen, E., Kristensen, G., Holm, R.,
241
Johansson, B., Hagmar, B., 1996. Use of multiple PCR primer sets for optimal
242
detection of human papillomavirus. J Clin Microbiol 34, 2095-2100.
M
240
Lee, J.K., Hong, J.H., Kang, S., Kim, D.Y., Kim, B.G., Kim, S.H., Kim, Y.M., Kim,
244
J.W., Kim, J.H., Kim, T.J., et al., 2013. Practice guidelines for the early detection
245
of cervical cancer in Korea: Korean Society of Gynecologic Oncology and the
246
Korean Society for Cytopathology 2012 edition. J Gynecol Oncol 24, 186-203.
ce pt
ed
243
Meijer, C.J., Berkhof, J., Castle, P.E., Hesselink, A.T., Franco, E.L., Ronco, G., Arbyn,
248
M., Bosch, F.X., Cuzick, J., Dillner, J., et al., 2009. Guidelines for human
249
papillomavirus DNA test requirements for primary cervical cancer screening in
250
Ac
247
women 30 years and older. Int J Cancer 124, 516-520.
251
Munoz, N., Bosch, F.X., de Sanjose, S., Herrero, R., Castellsague, X., Shah, K.V.,
252
Snijders, P.J., Meijer, C.J., 2003. Epidemiologic classification of human
253
papillomavirus types associated with cervical cancer. N Engl J Med 348, 518-527.
254
Poljak, M., Kovanda, A., Kocjan, B.J., Seme, K., Jancar, N., Vrtacnik-Bokal, E., 2009.
255
The Abbott RealTime High Risk HPV test: comparative evaluation of analytical 12
Page 16 of 17
256
specificity and clinical sensitivity for cervical carcinoma and CIN 3 lesions with
257
the Hybrid Capture 2 HPV DNA test. Acta Dermatovenerol Alp Panonica Adriat
258
18, 94-103. Poljak, M., Kocjan, B.J., 2010. Commercially available assays for multiplex detection
260
of alpha human papillomaviruses. Expert Rev Anti Infect Ther 8, 1139-1162.
261
Poljak, M., Ostrbenk, A., Seme, K., Ucakar, V., Hillemanns, P., Bokal, E.V., Jancar, N.,
262
Klavs, I., 2011. Comparison of clinical and analytical performance of the Abbott
263
Realtime High Risk HPV test to the performance of hybrid capture 2 in
264
population-based cervical cancer screening. J Clin Microbiol 49, 1721-1729.
265
Saslow, D., Solomon, D., Lawson, H.W., Killackey, M., Kulasingam, S.L., Cain, J.M.,
266
Garcia, F.A., Moriarty, A.T., Waxman, A.G., Wilbur, D.C., et al., 2012. American
267
Cancer Society, American Society for Colposcopy and Cervical Pathology, and
268
American Society for Clinical Pathology screening guidelines for the prevention
269
and early detection of cervical cancer. J Low Genit Tract Dis 16, 175-204.
ed
M
an
us
cr
ip t
259
Smith, J.S., Lindsay, L., Hoots, B., Keys, J., Franceschi, S., Winer, R., Clifford, G.M.,
271
2007. Human papillomavirus type distribution in invasive cervical cancer and
272
high-grade cervical lesions: a meta-analysis update. Int J Cancer 121, 621-632.
ce pt
270
Tang, N., Huang, S., Erickson, B., Mak, W.B., Salituro, J., Robinson, J., Abravaya, K.,
274
2009. High-risk HPV detection and concurrent HPV 16 and 18 typing with Abbott
275
RealTime High Risk HPV test. J Clin Virol 45 Suppl 1, S25-28.
Ac
273
13
Page 17 of 17