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Prevalence of resistance associated mutations in treatment-experienced HIV infected individuals in Latvia
Abstracts: Posters IP'1(341 Human immunodeficiency virus (HIV) sero-conversion in pregnancy - a case report S. Skidmore 1 , J. Weaver 1 , E. Smit 2. 1The Shrewsbury and
Telford Hospital NHS Trust, UK, 2HPA, Birmingham, Public Health Laboratory, Heart of England HNS Trust, UK Management of HIV infection during pregnancy can reduce mother to baby transmission to less than 1% and for this reason anti-HIV testing was introduced as part of routine antenatal care in England in 2000. However, since screening is carried out on a sample collected at around 12 weeks into the pregnancy, there is a potential for infection to occur after this time if there is a continuing risk. A 3-week old baby boy was admitted to hospital with signs of thrombocytopaenia and pneumonia. A urine sample was found to be CMV PCR positive with a CMV plasma viral load of 1.7 x 107 copies/ml. The clinical response to ganciclovir was a poor and various causes of immuno-suppression were considered. He was found to be HIV positive with a high viral load and a CD4 count of 20cells/mm3. HAART was instigated which resulted in a clinical improvement. The mother's booking blood was re-tested and confirmed negative but both the mother and father, on retesting at this time, were found to be HIV-positive. There was no sero-conversion type illness reported during pregnancy although incidence assay results were consistent with infection in the previous 3-6 months. Our case study exposes a weakness of the current screening programme and emphasizes the need for strict observance of universal precautions during labour and identification of mothers at risk of HIV infection. Babies infected in utero can have a rapid clinical progression and HIV should be part of the differential diagnosis in severely ill babies.
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L. Guseva, D. Dusacka, T. Kolupajeva, J. Storozenko, G. Sture, B. Rozentale. Infectology Center of Latvia, Riga, Latvia Aim of this study was to evaluate prevalence of HIV drug resistance associated mutations (RAM) in treatment-experienced patients in Latvia and to analyze drug susceptibility prediction data. Methods: Drug resistance testing was performed in 34 HIV-1 infected treatment-experienced individuals with treatment failures (20 male, treatment course from 1 till 15 years). Commercial kits VERSANT HIV-1 (LiPA) as well TRUGENE HIV-1 genotyping assay (BayerHealthCare-diagnostics) were used. Drug susceptibility was predicted using kits expert interpretations. Results: 30 (88%) out of 34 treatment-experienced patients had at least one RAM. Most frequently RAM were found for NRTI (23/34, 68%) followed by PI (21/34, 62%) and NNRTI (4/34, 12%). In the group of NRTI mutations more frequently occurred M184V (12/34), M41L (7/34) and T215Y (7/34), in PI mutations - major V82A (10/34), L90M (8/34), M461 (5/34) and minor M361/V (15/34), L89M (13/34), H69K (11/34). Most frequent NNRTI mutation was K103N (4/34). RAM resulted to the following drugs susceptibility: in 14 (41.2%) patients no evidence of resistance were found, in 4 (11. 8%) patients resistance to 1 drug class (NRTI or PI), in 12 patients (35.2%) resistance to 2 drugs classes (NRTI + PI) and in 4 (11.8%) patients resistance to all 3 classes of drugs (NRTI+NNRTI +PI) were predicted. Conclusions: In treated HIV infected individuals extremely high prevalence of RAM was found in the study on the limited number of specimen. In more then half cases resistance to at least one drug class was predicted. Resistance to 2 drug classes NRTI and PI was more typical. 1 ~
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NucliSens | easyMAG AND NucliSens | easyQ HIV-1 combination exhibits excellent performance characteristics
A. Buiting 1 , T. Oosterlaken 2, M. de Kock 2, E Simons 2, I. Berghuis 2.
1St. Elisabeth Hospital, Tilburg, The Netherlands, 2bioM6rieux, R&D Molecular Diagnostics, Boxtel, The Netherlands Background: Measurement of HIV-1 viral load is an accepted method to monitor the course of disease and the response to antiretroviral therapy in HIV-1 infected patients. The NucliSens EasyQ HIV-1 assay is an established viral load determination method, which is based on NASBA amplification and real-time detection using molecular beacon probes. Recently, two new front-end systems for magnetic nucleic acid extraction were developed: the NucliSens miniMAG and the NucliSens easyMAG. Method: Nucleic acids from analytical and subtype panels were isolated using the NucliSens miniMAG and easyMAG. HIV-1 RNA was amplified using the NucliSens EasyQ HIV-1 assay and amplicon formation was monitored using the NucliSens EasyQ Analyzer. Assay performance was assessed by the statistical analyses of detection limit, quantitative range, subtype detection and diagnostic specificity. Results: Analyses of an analytical panel with NucliSens EasyQ HIV-1 demonstrated accurate HIV-1 viral load monitoring over a range of 161-3x1061U/ml with a detection limit of 731U/ml. Analyses of FDA and BBI HIV-1 subtype panels showed successful detection of subtypes A through J and linearity was confirmed. Analyses of 114 HIV-1 negative donor samples demonstrated a 100% diagnostic specificity. Conclusion: The combination of NucliSens miniMAG or easyMAG with NucliSens EasyQ HIV-1 showed to be efficient and reliable. HIV-1 viral load monitoring is subtype independent and quantitative over a broad range. While NucliSens miniMAG extraction provides a cost efficient solution for low or medium throughput laboratories, NucliSens easyMAG extraction is optimally suited for high volume labs by offering high throughput and high level of user convenience.
Prevalence of resistance associated mutations in treatment-experienced HIV infected individuals in Latvia
Aseptic meningitis as a first sign of acute HIV infection
M. Bruckov~ 1, V. Chmelik 2, M. Linka 1, J. Vandasov~ 1, J. Schramlov~ 2, L. Houserov~ 2, Z. Jerhotov~ 2. 1National Reference
Laboratory on AIDS, Nail. Inst. of Publ. Hlth., Prague, Czech Republic, 2Dpt. of Infectious Diseases, AIDS Centre, Hospital Ceske Budejovice, Czech Republic Background: Although the clinical picture of HIV/AIDS is highly variable, HIV etiology may not be suspected in patients with atypical signs and symptoms and no history of HIV exposure. A HIV case with an unusual clinical presentation is reported. Methods: In 2004, a 20-year-old woman with high fever, signs of dysuria and meningeal irritation was admitted to the department of infectious diseases of the hospital after unsuccessful treatment at the departments of surgery, gynecology and urology. Based on analysis of cerebrospinal fluid (CSF), aseptic meningitis was diagnosed. With the patient's consent, plasma and CSF were screened for HIV. Quantitative RNA PCR assays for HIV in plasma and CSF, HIV antibodies in CSF, HIV genotyping and CD4 count were performed. Viral particles in CSF and plasma were detected by electron microscopy (ELM). Results: A patient diagnosed with aseptic meningitis tested positive for HIV antibodies in serum, plasma and CSR A low CD4 count (340/mm3) supported HIV diagnosis. An extremely high HIV-1 RNA viral load (5.270,000 RNAcopies/ml CFS) was found in CSE ELM detected picornaviral and retroviral particles. Diagnosis of aseptic meningitis due to HIV and opportunistic infection with picornaviruses was made. HAART successfully eradicated HIV from the CSF and plasma. Clinical improvement was obtained with specific ART. Conclusion: As clinical signs and symptoms of HIV infection vary widely, they should be suspected even in patients with unusual clinical patterns and no apparent history of HIV exposure. The reluctance to tell the truth about sexual habits and behaviour should be taken into account to avoid misdiagnosis.
Telford Hospital NHS Trust, UK, 2HPA, Birmingham, Public Health Laboratory, Heart of England HNS Trust, UK Management of HIV infection during pregnancy can reduce mother to baby transmission to less than 1% and for this reason anti-HIV testing was introduced as part of routine antenatal care in England in 2000. However, since screening is carried out on a sample collected at around 12 weeks into the pregnancy, there is a potential for infection to occur after this time if there is a continuing risk. A 3-week old baby boy was admitted to hospital with signs of thrombocytopaenia and pneumonia. A urine sample was found to be CMV PCR positive with a CMV plasma viral load of 1.7 x 107 copies/ml. The clinical response to ganciclovir was a poor and various causes of immuno-suppression were considered. He was found to be HIV positive with a high viral load and a CD4 count of 20cells/mm3. HAART was instigated which resulted in a clinical improvement. The mother's booking blood was re-tested and confirmed negative but both the mother and father, on retesting at this time, were found to be HIV-positive. There was no sero-conversion type illness reported during pregnancy although incidence assay results were consistent with infection in the previous 3-6 months. Our case study exposes a weakness of the current screening programme and emphasizes the need for strict observance of universal precautions during labour and identification of mothers at risk of HIV infection. Babies infected in utero can have a rapid clinical progression and HIV should be part of the differential diagnosis in severely ill babies.
$53
1•
L. Guseva, D. Dusacka, T. Kolupajeva, J. Storozenko, G. Sture, B. Rozentale. Infectology Center of Latvia, Riga, Latvia Aim of this study was to evaluate prevalence of HIV drug resistance associated mutations (RAM) in treatment-experienced patients in Latvia and to analyze drug susceptibility prediction data. Methods: Drug resistance testing was performed in 34 HIV-1 infected treatment-experienced individuals with treatment failures (20 male, treatment course from 1 till 15 years). Commercial kits VERSANT HIV-1 (LiPA) as well TRUGENE HIV-1 genotyping assay (BayerHealthCare-diagnostics) were used. Drug susceptibility was predicted using kits expert interpretations. Results: 30 (88%) out of 34 treatment-experienced patients had at least one RAM. Most frequently RAM were found for NRTI (23/34, 68%) followed by PI (21/34, 62%) and NNRTI (4/34, 12%). In the group of NRTI mutations more frequently occurred M184V (12/34), M41L (7/34) and T215Y (7/34), in PI mutations - major V82A (10/34), L90M (8/34), M461 (5/34) and minor M361/V (15/34), L89M (13/34), H69K (11/34). Most frequent NNRTI mutation was K103N (4/34). RAM resulted to the following drugs susceptibility: in 14 (41.2%) patients no evidence of resistance were found, in 4 (11. 8%) patients resistance to 1 drug class (NRTI or PI), in 12 patients (35.2%) resistance to 2 drugs classes (NRTI + PI) and in 4 (11.8%) patients resistance to all 3 classes of drugs (NRTI+NNRTI +PI) were predicted. Conclusions: In treated HIV infected individuals extremely high prevalence of RAM was found in the study on the limited number of specimen. In more then half cases resistance to at least one drug class was predicted. Resistance to 2 drug classes NRTI and PI was more typical. 1 ~
I•
NucliSens | easyMAG AND NucliSens | easyQ HIV-1 combination exhibits excellent performance characteristics
A. Buiting 1 , T. Oosterlaken 2, M. de Kock 2, E Simons 2, I. Berghuis 2.
1St. Elisabeth Hospital, Tilburg, The Netherlands, 2bioM6rieux, R&D Molecular Diagnostics, Boxtel, The Netherlands Background: Measurement of HIV-1 viral load is an accepted method to monitor the course of disease and the response to antiretroviral therapy in HIV-1 infected patients. The NucliSens EasyQ HIV-1 assay is an established viral load determination method, which is based on NASBA amplification and real-time detection using molecular beacon probes. Recently, two new front-end systems for magnetic nucleic acid extraction were developed: the NucliSens miniMAG and the NucliSens easyMAG. Method: Nucleic acids from analytical and subtype panels were isolated using the NucliSens miniMAG and easyMAG. HIV-1 RNA was amplified using the NucliSens EasyQ HIV-1 assay and amplicon formation was monitored using the NucliSens EasyQ Analyzer. Assay performance was assessed by the statistical analyses of detection limit, quantitative range, subtype detection and diagnostic specificity. Results: Analyses of an analytical panel with NucliSens EasyQ HIV-1 demonstrated accurate HIV-1 viral load monitoring over a range of 161-3x1061U/ml with a detection limit of 731U/ml. Analyses of FDA and BBI HIV-1 subtype panels showed successful detection of subtypes A through J and linearity was confirmed. Analyses of 114 HIV-1 negative donor samples demonstrated a 100% diagnostic specificity. Conclusion: The combination of NucliSens miniMAG or easyMAG with NucliSens EasyQ HIV-1 showed to be efficient and reliable. HIV-1 viral load monitoring is subtype independent and quantitative over a broad range. While NucliSens miniMAG extraction provides a cost efficient solution for low or medium throughput laboratories, NucliSens easyMAG extraction is optimally suited for high volume labs by offering high throughput and high level of user convenience.
Prevalence of resistance associated mutations in treatment-experienced HIV infected individuals in Latvia
Aseptic meningitis as a first sign of acute HIV infection
M. Bruckov~ 1, V. Chmelik 2, M. Linka 1, J. Vandasov~ 1, J. Schramlov~ 2, L. Houserov~ 2, Z. Jerhotov~ 2. 1National Reference
Laboratory on AIDS, Nail. Inst. of Publ. Hlth., Prague, Czech Republic, 2Dpt. of Infectious Diseases, AIDS Centre, Hospital Ceske Budejovice, Czech Republic Background: Although the clinical picture of HIV/AIDS is highly variable, HIV etiology may not be suspected in patients with atypical signs and symptoms and no history of HIV exposure. A HIV case with an unusual clinical presentation is reported. Methods: In 2004, a 20-year-old woman with high fever, signs of dysuria and meningeal irritation was admitted to the department of infectious diseases of the hospital after unsuccessful treatment at the departments of surgery, gynecology and urology. Based on analysis of cerebrospinal fluid (CSF), aseptic meningitis was diagnosed. With the patient's consent, plasma and CSF were screened for HIV. Quantitative RNA PCR assays for HIV in plasma and CSF, HIV antibodies in CSF, HIV genotyping and CD4 count were performed. Viral particles in CSF and plasma were detected by electron microscopy (ELM). Results: A patient diagnosed with aseptic meningitis tested positive for HIV antibodies in serum, plasma and CSR A low CD4 count (340/mm3) supported HIV diagnosis. An extremely high HIV-1 RNA viral load (5.270,000 RNAcopies/ml CFS) was found in CSE ELM detected picornaviral and retroviral particles. Diagnosis of aseptic meningitis due to HIV and opportunistic infection with picornaviruses was made. HAART successfully eradicated HIV from the CSF and plasma. Clinical improvement was obtained with specific ART. Conclusion: As clinical signs and symptoms of HIV infection vary widely, they should be suspected even in patients with unusual clinical patterns and no apparent history of HIV exposure. The reluctance to tell the truth about sexual habits and behaviour should be taken into account to avoid misdiagnosis.