- Email: [email protected]
Quick diagnosis of female genital tuberculosis using multiplex fast polymerase chain reaction in Southern Assam, India
72
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adaptive, prophylactic function, i.e. the maternal and embryo protection hypothesis, by avoiding foods that may contain harmful material and triggering aversions to these kind of foods during pregnancy [4].
Conflict of interest The authors have no conflicts of interest to declare.
References [1] Adam I, Khamis AH, Elbashir MI. Prevalence and risk factors for anaemia in pregnant women of eastern Sudan. Trans R Soc Trop Med Hyg 2005;99(10):739–43. [2] Sule S, Madugu HN. Pica in pregnant women in Zaria, Nigeria. Niger J Med 2001;10(1): 25–7. [3] Nyaruhucha CN. Food cravings, aversions and pica among pregnant women in Dar es Salaam, Tanzania. Tanzan J Health Res 2009;11(1):29–34. [4] Flaxman SM, Sherman PW. Morning sickness: a mechanism for protecting mother and embryo. Q Rev Biol 2000;75(2):113–48.
0020-7292/$ – see front matter © 2012 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.ijgo.2012.02.004
Quick diagnosis of female genital tuberculosis using multiplex fast polymerase chain reaction in Southern Assam, India Sankar Kumar Ghosh ⁎, Rosy Mondal Department of Biotechnology, Assam University, Silchar, Assam, India
a r t i c l e
i n f o
Article history: Received 5 December 2011 Received in revised form 1 February 2012 Accepted 12 March 2012 Keywords: Endometrial biopsy specimens Female genital tuberculosis India Menstrual blood Multiplex fast polymerase chain reaction
Tuberculosis-related subfertility causing tubal dysfunction is a major concern in India. Female genital tuberculosis is responsible for about 26% of the infertility among Indian women despite specific therapy [1,2]. The aim of the present study was to investigate the rapid and specific detection of Mycobacterium tuberculosis in infertile patients in Southern Assam, India. The study was conducted at Assam University, Silchar between May 2010 and April 2011. DNA was isolated from endometrial biopsy and menstrual blood specimens of 124 infertile women (20–40 years), 60 of whom were clinically suspected to have female genital tuberculosis. A 240-bp region of the MPT64 gene and a 541-bp region of the IS6110 gene were amplified using 2 sets of primers. Multiplex fast polymerase chain reaction (PCR) using the AmpliTaq Gold PCR master mix (Applied Biosystems, Carlsbad, CA, USA) was used for quick diagnosis (within 15 minutes) and compared with conventional PCR and routine cytological investigation [3]. The number of positive cases identified using endometrial and menstrual blood specimens was 36.7% (22 out of 60) using cytological analysis compared with 43.3% (26 out of 60) with PCR amplification. When PCR was carried out using the menstrual blood specimens of the 64 infertile patients who were not suspected to
⁎ Corresponding author at: Department of Biotechnology, Assam University, Silchar, Pin 788011, Assam, India. Tel.: +91 9435 372338; fax: +91 3842 270806. E-mail address: [email protected] (S.K. Ghosh).
have female genital tuberculosis, 18.7% (12 out of 64) of samples were positive. This was later confirmed by collecting endometrial tissue from the same patients. Multiplex PCR using 2 sets of primers to produce 2 distinct bands on gel eliminates the possibility of false positivity by PCR methods (Fig. 1). As a result of PCR detection, 2 infertile women conceived following specific treatment for tuberculosis, which went undetected by cytological staining. Endometrial biopsy is the universal standard for detection of female genital tuberculosis. In the present study, PCR of the target gene in both endometrial biopsy and menstrual blood specimens showed consistent results. Both conventional multiplex PCR and fast PCR were conducted to validate the results since PCR is more sensitive than routine cytological methods [4]. An effective and accurate method of diagnosing female genital tuberculosis should be the combination of PCR and traditional cytological analysis. Multiplex fast PCR is rapid, sensitive, and specific in detecting asymptomatic female genital tuberculosis, thereby facilitating early therapeutic decisions in suspected cases. Acknowledgments The project is funded by the Department of Biotechnology, Government of India. Conflict of interest The authors have no conflicts of interest to declare. References [1] Gupta N, Sharma JB, Mittal S, Singh N, Misra R, Kukreja M. Genital tuberculosis in Indian infertility patients. Int J Gynecol Obstet 2007;97(2):135–8. [2] Agashe V, Shenai S, Mohrir G, Deshmukh M, Bhaduri A, Deshpande R, et al. Osteoarticular tuberculosis–diagnostic solutions in a disease endemic region. J Infect Dev Ctries 2009;3(7):511–6. [3] Ghosh SK, Choudhury B, Hansa J, Mondal R, Singh M, Duttagupta S, et al. Human papillomavirus testing for suspected cervical cancer patients from Southern Assam by fast-PCR. Asian Pac J Cancer Prev 2011;12(3):749–51. [4] Qin L, Zheng R, Fan C, Cai J, Liu Z, Wang J, et al. Identification and evaluation of a new nucleic acid amplification test target for specific detection of Mycobacterium tuberculosis. Clin Chem Lab Med 2010;48(10):1501–5.
BRIEF COMMUNICATIONS
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Fig. 1. PCR amplified products of MPT64 and IS6110 genes on 1.5% agarose gel. (A) Conventional PCR: B1–B6 represents menstrual blood and E1–E6 represents endometrial biopsy specimens; (B) Multiplex fast PCR: B1, B2, and B3 represent menstrual blood and E1, E2, and E3 represent endometrial biopsy specimens.
0020-7292/$ – see front matter © 2012 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.ijgo.2012.02.006
Effect of the gonadotropin-releasing hormone antagonist cetrorelix on the prevention of chemotherapy-induced ovarian damage in women with hematological malignancy Agathi Digeni a, Argiris Symeonidis b, Neoklis A. Georgopoulos a, c,⁎ a b c
Department of Obstetrics and Gynecology, University of Patras Medical School, Patras, Greece Department of Internal Medicine, Division of Hematology, University of Patras Medical School, Patras, Greece Division of Reproductive Endocrinology, University of Patras Medical School, Patras, Greece
a r t i c l e
i n f o
Article history: Received 12 December 2011 Received in revised form 16 February 2012 Accepted 16 March 2012 Keywords: Cetrorelix Chemotherapy Fertility GnRH antagonist Hematologic malignancies Ovarian damage
Gonadotropin-releasing hormone (GnRH) agonists are the most commonly used pharmaceutical treatment for ovarian protection during chemotherapy [1,2]. A recent study showed that a combination of GnRH agonists and antagonists induced a long-lasting suppression of gonadotropins but did not completely prevent flare-up of follicle stimulating hormone [3]. The aim of the present study was to evaluate the use of cetrorelix, a GnRH antagonist, for prevention of ovarian damage during chemotherapy in patients with a hematologic malignancy, such as Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), and acute leukemia (AL). The study included 24 women (mean age at diagnosis, 23.3 ± 6.4 years) who had been diagnosed with a hematologic malignancy ⁎ Corresponding author at: Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology, University of Patras Medical School, Rio-26500, Greece. Tel.: +30 2610 999835; fax: +30 2610 993854. E-mail address: [email protected] (N.A. Georgopoulos).
(15 HL, 4 NHL, and 5 AL). All patients received chemotherapy with various cytotoxic agents, including cyclophosphamide, and 3 mg of cetrorelix subcutaneously every second day. Data were compared retrospectively with 31 women (mean age, 26.4 ± 6.2 years) who had been diagnosed with a hematologic cancer (18 HL, 12 NHL, and 3 AL) and received the same chemotherapy, but without cetrorelix. The study was approved by the Institutional Review Board. P b 0.05 was considered significant at statistical analysis. Among the 23 evaluable women (1 died) who received cetrorelix, the incidence of secondary amenorrhea 6 months after cessation of treatment was significantly lower compared with the women who did not receive cetrorelix (4.3% vs 27.3%, P = 0.036; Table 1). No significant differences were observed between the 2 groups in the mean serum levels of follicle-stimulating hormone, estradiol, and anti-Müllerian hormone after completion of chemotherapy (P > 0.05; Table 1), suggesting a degree of ovarian damage despite cetrorelix administration.
Table 1 Clinical and hormonal data from women treated with cytotoxic agents for hematologic cancer, with or without cetrorelix administration for prevention of ovarian damage.a Variables
No ovarian protection (n = 33)
Cetrorelix GnRH antagonist (n = 23)
P value
Age at diagnosis, y Follicle-stimulating hormone, mIU/mL Estradiol, pg/mL Anti-Müllerian hormone, ng/mL Secondary amenorrhea
26.36 ± 6.19 20.91 ± 22.41 55.16 ± 32.04 0.39 ± 0.67 9 (27.3)
23.33 ± 6.45 22.17 ± 31.14 77.57 ± 53.96 0.38 ± 0.56 1 (4.3)
0.090 0.682 0.205 0.466 0.036
a
Values are given as mean ± SD or number (percentage).
BRIEF COMMUNICATIONS
adaptive, prophylactic function, i.e. the maternal and embryo protection hypothesis, by avoiding foods that may contain harmful material and triggering aversions to these kind of foods during pregnancy [4].
Conflict of interest The authors have no conflicts of interest to declare.
References [1] Adam I, Khamis AH, Elbashir MI. Prevalence and risk factors for anaemia in pregnant women of eastern Sudan. Trans R Soc Trop Med Hyg 2005;99(10):739–43. [2] Sule S, Madugu HN. Pica in pregnant women in Zaria, Nigeria. Niger J Med 2001;10(1): 25–7. [3] Nyaruhucha CN. Food cravings, aversions and pica among pregnant women in Dar es Salaam, Tanzania. Tanzan J Health Res 2009;11(1):29–34. [4] Flaxman SM, Sherman PW. Morning sickness: a mechanism for protecting mother and embryo. Q Rev Biol 2000;75(2):113–48.
0020-7292/$ – see front matter © 2012 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.ijgo.2012.02.004
Quick diagnosis of female genital tuberculosis using multiplex fast polymerase chain reaction in Southern Assam, India Sankar Kumar Ghosh ⁎, Rosy Mondal Department of Biotechnology, Assam University, Silchar, Assam, India
a r t i c l e
i n f o
Article history: Received 5 December 2011 Received in revised form 1 February 2012 Accepted 12 March 2012 Keywords: Endometrial biopsy specimens Female genital tuberculosis India Menstrual blood Multiplex fast polymerase chain reaction
Tuberculosis-related subfertility causing tubal dysfunction is a major concern in India. Female genital tuberculosis is responsible for about 26% of the infertility among Indian women despite specific therapy [1,2]. The aim of the present study was to investigate the rapid and specific detection of Mycobacterium tuberculosis in infertile patients in Southern Assam, India. The study was conducted at Assam University, Silchar between May 2010 and April 2011. DNA was isolated from endometrial biopsy and menstrual blood specimens of 124 infertile women (20–40 years), 60 of whom were clinically suspected to have female genital tuberculosis. A 240-bp region of the MPT64 gene and a 541-bp region of the IS6110 gene were amplified using 2 sets of primers. Multiplex fast polymerase chain reaction (PCR) using the AmpliTaq Gold PCR master mix (Applied Biosystems, Carlsbad, CA, USA) was used for quick diagnosis (within 15 minutes) and compared with conventional PCR and routine cytological investigation [3]. The number of positive cases identified using endometrial and menstrual blood specimens was 36.7% (22 out of 60) using cytological analysis compared with 43.3% (26 out of 60) with PCR amplification. When PCR was carried out using the menstrual blood specimens of the 64 infertile patients who were not suspected to
⁎ Corresponding author at: Department of Biotechnology, Assam University, Silchar, Pin 788011, Assam, India. Tel.: +91 9435 372338; fax: +91 3842 270806. E-mail address: [email protected] (S.K. Ghosh).
have female genital tuberculosis, 18.7% (12 out of 64) of samples were positive. This was later confirmed by collecting endometrial tissue from the same patients. Multiplex PCR using 2 sets of primers to produce 2 distinct bands on gel eliminates the possibility of false positivity by PCR methods (Fig. 1). As a result of PCR detection, 2 infertile women conceived following specific treatment for tuberculosis, which went undetected by cytological staining. Endometrial biopsy is the universal standard for detection of female genital tuberculosis. In the present study, PCR of the target gene in both endometrial biopsy and menstrual blood specimens showed consistent results. Both conventional multiplex PCR and fast PCR were conducted to validate the results since PCR is more sensitive than routine cytological methods [4]. An effective and accurate method of diagnosing female genital tuberculosis should be the combination of PCR and traditional cytological analysis. Multiplex fast PCR is rapid, sensitive, and specific in detecting asymptomatic female genital tuberculosis, thereby facilitating early therapeutic decisions in suspected cases. Acknowledgments The project is funded by the Department of Biotechnology, Government of India. Conflict of interest The authors have no conflicts of interest to declare. References [1] Gupta N, Sharma JB, Mittal S, Singh N, Misra R, Kukreja M. Genital tuberculosis in Indian infertility patients. Int J Gynecol Obstet 2007;97(2):135–8. [2] Agashe V, Shenai S, Mohrir G, Deshmukh M, Bhaduri A, Deshpande R, et al. Osteoarticular tuberculosis–diagnostic solutions in a disease endemic region. J Infect Dev Ctries 2009;3(7):511–6. [3] Ghosh SK, Choudhury B, Hansa J, Mondal R, Singh M, Duttagupta S, et al. Human papillomavirus testing for suspected cervical cancer patients from Southern Assam by fast-PCR. Asian Pac J Cancer Prev 2011;12(3):749–51. [4] Qin L, Zheng R, Fan C, Cai J, Liu Z, Wang J, et al. Identification and evaluation of a new nucleic acid amplification test target for specific detection of Mycobacterium tuberculosis. Clin Chem Lab Med 2010;48(10):1501–5.
BRIEF COMMUNICATIONS
73
Fig. 1. PCR amplified products of MPT64 and IS6110 genes on 1.5% agarose gel. (A) Conventional PCR: B1–B6 represents menstrual blood and E1–E6 represents endometrial biopsy specimens; (B) Multiplex fast PCR: B1, B2, and B3 represent menstrual blood and E1, E2, and E3 represent endometrial biopsy specimens.
0020-7292/$ – see front matter © 2012 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.ijgo.2012.02.006
Effect of the gonadotropin-releasing hormone antagonist cetrorelix on the prevention of chemotherapy-induced ovarian damage in women with hematological malignancy Agathi Digeni a, Argiris Symeonidis b, Neoklis A. Georgopoulos a, c,⁎ a b c
Department of Obstetrics and Gynecology, University of Patras Medical School, Patras, Greece Department of Internal Medicine, Division of Hematology, University of Patras Medical School, Patras, Greece Division of Reproductive Endocrinology, University of Patras Medical School, Patras, Greece
a r t i c l e
i n f o
Article history: Received 12 December 2011 Received in revised form 16 February 2012 Accepted 16 March 2012 Keywords: Cetrorelix Chemotherapy Fertility GnRH antagonist Hematologic malignancies Ovarian damage
Gonadotropin-releasing hormone (GnRH) agonists are the most commonly used pharmaceutical treatment for ovarian protection during chemotherapy [1,2]. A recent study showed that a combination of GnRH agonists and antagonists induced a long-lasting suppression of gonadotropins but did not completely prevent flare-up of follicle stimulating hormone [3]. The aim of the present study was to evaluate the use of cetrorelix, a GnRH antagonist, for prevention of ovarian damage during chemotherapy in patients with a hematologic malignancy, such as Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), and acute leukemia (AL). The study included 24 women (mean age at diagnosis, 23.3 ± 6.4 years) who had been diagnosed with a hematologic malignancy ⁎ Corresponding author at: Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology, University of Patras Medical School, Rio-26500, Greece. Tel.: +30 2610 999835; fax: +30 2610 993854. E-mail address: [email protected] (N.A. Georgopoulos).
(15 HL, 4 NHL, and 5 AL). All patients received chemotherapy with various cytotoxic agents, including cyclophosphamide, and 3 mg of cetrorelix subcutaneously every second day. Data were compared retrospectively with 31 women (mean age, 26.4 ± 6.2 years) who had been diagnosed with a hematologic cancer (18 HL, 12 NHL, and 3 AL) and received the same chemotherapy, but without cetrorelix. The study was approved by the Institutional Review Board. P b 0.05 was considered significant at statistical analysis. Among the 23 evaluable women (1 died) who received cetrorelix, the incidence of secondary amenorrhea 6 months after cessation of treatment was significantly lower compared with the women who did not receive cetrorelix (4.3% vs 27.3%, P = 0.036; Table 1). No significant differences were observed between the 2 groups in the mean serum levels of follicle-stimulating hormone, estradiol, and anti-Müllerian hormone after completion of chemotherapy (P > 0.05; Table 1), suggesting a degree of ovarian damage despite cetrorelix administration.
Table 1 Clinical and hormonal data from women treated with cytotoxic agents for hematologic cancer, with or without cetrorelix administration for prevention of ovarian damage.a Variables
No ovarian protection (n = 33)
Cetrorelix GnRH antagonist (n = 23)
P value
Age at diagnosis, y Follicle-stimulating hormone, mIU/mL Estradiol, pg/mL Anti-Müllerian hormone, ng/mL Secondary amenorrhea
26.36 ± 6.19 20.91 ± 22.41 55.16 ± 32.04 0.39 ± 0.67 9 (27.3)
23.33 ± 6.45 22.17 ± 31.14 77.57 ± 53.96 0.38 ± 0.56 1 (4.3)
0.090 0.682 0.205 0.466 0.036
a
Values are given as mean ± SD or number (percentage).