Epidemiology and Determinants of Serologically Diagnosed HIV-1 and HIV-1&2 in Tertiary Hospitals of Eastern Peninsular Malaysia

@#Introduction: HIV is the leading cause of mortality and morbidity worldwide. There are two types of HIV, HIV-1, and HIV-2, which are geographically different in epidemiology and determinants. Objective: To determine the epidemiology and determinants of HIV-1 and HIV-1&2 in the three tertiary hospitals of Eastern Peninsular Malaysia. Method: A cross-sectional study of confirmed serologically HIV-1 and HIV-1&2 from January 2016 until December 2018. SPSS analysed all collected data, descriptive statistics for sociodemographic data and Pearson chi-square for the association between type of HIV with HCV, HBV, syphilis, and tuberculosis. In identifying the risk factor associated with HIV-1&2, several variables were tested by the Multiple Logistic Regression Model. A P-value of <0.05 was considered statistically significant. Results: Out of 519 serologically diagnosed HIV, 344 (66.28%) were HIV-1, and 175 (33.72%) were HIV-1&2. HIV positive were highly distributed in Malay male in both groups. Most HIV-1 patients were single, unemployed, and presented with tuberculosis. HIV-1&2 were single and employed, mostly asymptomatic at diagnosis. The commonest mode of transmission for HIV-1 was by sexual contact (31.87%), whereas IVDU (13.63%) in HIV-1&2. Co-infection with tuberculosis (P=0.005) and HCV (P<0.001) were significantly higher in HIV-1 as compared to HIV-1&2. IVDU was a significant determinant to develop HIV-1&2 (Adjusted OR: 3.5, 95% CI=1.875-5.227, P<0.001). Conclusion: HIV-1&2 was high in this study. Patients with HIV-1&2 present in less severe symptoms compared to the patient with HIV-1. A further molecular diagnostic study should be tested to confirm the type of HIV.

HIV-2 Treatment in Mali and Drug Resistance Antiretroviral Profile

Aims:We evaluated different treatment regimens administered to patients infected with HIV-2 in Mali, and studied pol polymorphisms that may influence susceptibility to antiretrovirals.Methodology:We collected 57 blood samples from HIV -2 seropositive patients in health centers in Mali (Bamako, Segou and Sikasso), including 21 treated patients and 36 untreated patients. The confirmation of the serological status and the measurement of the viral load were carried out in Brussels. Genotypic analysis of protease, reverse transcriptase and integrase was then performed on specimens with detectable viral load. The search for ARV resistance mutations and polymorphism positions was performed in comparison with reference sequences.Results:The most used treatment regimen was ZDV-3TC-LPV/ r. Genotypic analysis of protease, reverse transcriptase and integrase was performed on 20 samples from patients with detectable viral load: 16 untreated patients and 4 treated patients. Seventy-five percent of the strains analyzed correspond to group A of HIV-2. In 2cas, mutations associated with resistance to the administered molecules were found. Three viral strains from untreated patients had NRTI or PI resistance mutations. In integrase, no mutation associated with resistance was observed.Conclusion:The circulation of strains carrying resistance mutations to NRTIs and PIs has been observed in Mali

Progress in HIV-2 detection methodology / 中华流行病学杂志

Laboratory test is the routine method of diagnosis, monitoring and blood screening of HIV infection, and main basis for early diagnosis of AIDS. HIV is divided into HIV-1 and HIV-2 subtypes, HIV-1 infection is the major cause of AIDS pandemic, while HIV-2 infection occurs in limited areas in the world, mainly in West Africa. HIV-2 infection has been reported in China since 1998. They are sporadic cases, and mainly HIV-1/HIV-2 mixed infections. There are less concerns about HIV-2 detection in China at present, and domestic HIV-2 detection reagents have not come into the market. At present, the detection method of HIV-2 is mainly antibody test and nucleic acid test. The initial screening is through rapid test and other methods and the confirmation is depended on Western Blot and Line Immune Assay. According to the HIV antibody test results, HIV-2 infection is confirmed. With the rapid development of molecular biology, the diagnostic method of nucleic acid detection laboratory has made great progress.

Diagnostic and epidemiological features of the first two HIV-2 indigenous infections in Hunan province / 中华流行病学杂志

Objective To study the diagnostic and epidemiological features of the first two HIV-2 indigenous cases in Hunan province.Methods Blood samples from two individuals with "HIV antibody indeterminate" and HIV-2 specific band showed by HIV-1/2 western blotting method,were repeatedly collected and detected under HIV 1 + 2 strip immunoassay and PCR,in Changsha city,Hunan province,through March to November,2017.An epidemiological survey was carried out at the same time.Results Our findings showed that the two cases were sex partners,without histories of sexual contact with foreigners and the source of infection was unknown.Results from the HIV 1 + 2 antibody confirmation test showed that they were "HIV-2 antibody positive".Through amplifying and sequencing the gag area of HIV-2 and BLAST,the similarity of HIV-2 strains presented as 98％.The results also showed that there were HIV-2 specific fragments in the two cases.Conclusion HIV-2 indigenous cases had never been reported in China.These cases had brought new challenge on prevention,diagnosis and treatment of HIV/AIDS in China.

Diagnostic and epidemiological features of the first two HIV-2 indigenous infections in Hunan province / 中华流行病学杂志

Objective To study the diagnostic and epidemiological features of the first two HIV-2 indigenous cases in Hunan province.Methods Blood samples from two individuals with "HIV antibody indeterminate" and HIV-2 specific band showed by HIV-1/2 western blotting method,were repeatedly collected and detected under HIV 1 + 2 strip immunoassay and PCR,in Changsha city,Hunan province,through March to November,2017.An epidemiological survey was carried out at the same time.Results Our findings showed that the two cases were sex partners,without histories of sexual contact with foreigners and the source of infection was unknown.Results from the HIV 1 + 2 antibody confirmation test showed that they were "HIV-2 antibody positive".Through amplifying and sequencing the gag area of HIV-2 and BLAST,the similarity of HIV-2 strains presented as 98％.The results also showed that there were HIV-2 specific fragments in the two cases.Conclusion HIV-2 indigenous cases had never been reported in China.These cases had brought new challenge on prevention,diagnosis and treatment of HIV/AIDS in China.

Evaluación de la estabilidad del sistema de diagnóstico inmunoenzimático DAVIH VIH-2 / Evaluation of the stability of the immunoenzymatic diagnostic system DAVIH VIH-2

Introducción: el estudio de la estabilidad de los componentes y el producto terminado constituye un importante requisito regulatorio en los diagnosticadores. Objetivo: realizar un estudio de estabilidad en tiempo real durante doce meses del sistema inmunoenzimático (ELISA) DAVIH VIH-2. Métodos: se realizó un estudio de estabilidad en tiempo real durante doce meses en tres lotes del diagnosticador DAVIH VIH-2, ELISA indirecto diseñado para la detección de anticuerpos contra el virus de inmunodeficiencia humana tipo 2 en suero o plasma humano. Se controlaron los requisitos de calidad de los componentes de acuerdo a sus especificaciones. Se estudió la normalidad de valores de densidad óptica/valor límite y la homogeneidad de las medias y varianzas mediante las dócimas de Grubbs y Cochran. Se estimó la precisión en los controles positivo y negativo del sistema y en seis muestras con diferente reactividad al virus de inmunodeficiencia humana tipo 2 mediante el cálculo del coeficiente de variación y se confeccionaron las cartas de control de los valores de las medias de densidad óptica respecto al tiempo. Resultados: los requisitos de calidad de cada componente se cumplieron durante 12 meses, excepto las características funcionales del conjugado a partir de los seis meses. Los valores en las dócimas de Grubbs y Cochran fueron menores que los valores críticos tabulados para α del 1 y 5 por ciento por lo que existió homogeneidad en las medias y las varianzas en todo el periodo. El coeficiente de variación se mantuvo inferior al 10 por ciento excepto en las muestras con reactividad media y baja, mientras que en las cartas de control, los valores de densidad óptica se mantuvieron en el límite de la media ±2 desviaciones estándar hasta el noveno mes(AU)

Seroprevalence of HIV-2 in and around Nagpur.

Abstracts: Backround: Human immunodeficiency virus type 2 (HIV- 2) belongs to the family Retroviridae and is morphologically similar to HIV-1. Reliable and up-to- date information on the HIV-2 epidemic in India is still lacking. Methodology: We conducted this study to know the seroprevalence of HIV-2 in our region. Results: A total 15046 samples were screened at ICTC that includes 6343 from ANC mothers and 8703 samples from direct walk-in clients and referred patients.for HIV-2 antibodies. Also, we found 9 (0.10%) samples positive for HIV-2. One patient was coinfected with HIV-1 and HIV-2. There were no HIV-2 positive cases in ANC and paediatric age group. Conclusion: As HIV-2 is being reported from various parts of the country and its treatment modalities differ from HIV-1 hence screening for HIV-2 should be carried out routinely.

Prevalence of HIV among blood donors in a tertiary care centre of north India.

Background & objectives: India has the second highest HIV population in the world with about 2.5-3.0 million cases. HIV-2 cases among general and blood donor population have also been reported mostly from west and south India. This single centre study was carried out to observe the HIV-1 and HIV-2 prevalence among blood donors from north India. Methods: A total of 2,04,677 people were screened for the presence of HIV infection over the 11 year period (1999 to 2009). Till 2004, a third generation ELISA kit was used. From 2005 till January 2009 all tests were done using the fourth generation ELISA kit which detected the presence of HIV-1 P24 antigen and anti-HIV antibodies. From February 2009 onwards, the kits used were Genscreen ULTRA HIV Ag- Ab Assay. Results: A total of 506 (0.247%) donors were found to be repeat reactive for HIV. Of these, 486 (96%) donors tested using the Western blot were found positive for HIV-1 infection. Twenty (4%) donors showed a negative Western blot result, none of the donors were found reactive for HIV-2 infection. Interpretation & conclusions: The prevalence of HIV was 0.249 per cent among blood donors of north India. No HIV-2 case was found among the studied blood donor population indicating that it is not a threat currently.

Evaluación de la inmovilización de un péptido de gp36 de VIH-2 en membranas de nitrocelulosa / Evaluation of the immobilization of one HIV-2 gp36 peptide in nitrocellulose membrane

Introducción: la inmovilización de antígenos a soportes sólidos se utiliza para el desarrollo de diversos inmunoensayos. Una de las primeras tecnologías desarrolladas fue la adsorción de proteínas por aplicación directa sobre la nitrocelulosa. Objetivo: normalizar la inmovilización de un péptido sintético de la proteína de transmembrana gp36 del VIH-2, a un soporte de nitrocelulosa para fines diagnósticos y evaluar los parámetros de desempeño en un grupo de muestras de sueros con reactividad de interés conocida. Métodos: el péptido se inmovilizó de forma libre, conjugado a la albúmina de suero bovina (BSA) y a la hemocianina de lapa marina (KLH) como proteínas portadoras. Se analizaron los parámetros de inmovilización y se determinó la variante óptima. Con la variante escogida se evaluó la sensibilidad y especificidad diagnóstica frente a paneles de referencia del Laboratorio de Investigaciones del SIDA. La especificidad analítica se evaluó con muestras reactivas a VIH-1 y HTLV-I. Resultados: el análisis de las variantes de péptido inmovilizadas a las membranas de nitrocelulosa, demostró que el péptido gp36-BSA, fue el que logró la mayor diferenciación entre muestras positivas y negativas. Se obtuvo 100 por ciento de sensibilidad y 95,2 por ciento de especificidad diagnóstica, así como 100 por ciento de especificidad analítica. Conclusiones: el péptido gp36-BSA inmovilizado en membranas de nitrocelulosa es eficaz en el diagnóstico serológico del VIH-2, lo cual permitirá considerarlo para su empleo con fines diagnósticos en sistemas que utilicen como fase sólida la nitrocelulosa.

Introduction: antigen immobilization in solid supports is used for the development of several immunoassays. One of the first technologies developed was the protein adsorption by direct application to nitrocellulose. Objective: to standardize the immobilization of a synthetic peptide of the HIV-2 transmembrane protein gp36 to nitrocellulose support for diagnostic purposes and to evaluate the performance parameters in a group of serum samples with recognized interesting reactivity. Methods: the peptide was freely immobilized, conjugated to bovine serum albumin (BSA) and to keyhole limpet hemocyanin (KLH) as carrier proteins. Immobilization parameters were analyzed and then, the optimal immobilization alternative was determined. Using the chosen variant, the diagnostic sensitivity and specificity against reference panels of the AIDS Research Laboratory were evaluated. Analytical specificity was evaluated with reactive samples to HIV-1 and HTLV-I. Results: the analysis of the immobilized peptide variants to nitrocellulose membranes showed that the gp36 peptide-BSA was the one that succeeded in setting the greatest differentiation between positive and negative samples. There were observed 100 percent sensitivity, 95.2 percent diagnostic specificity and 100 percent analytical specificity. Conclusions: the gp36-BSA peptide immobilized on nitrocellulose membranes showed efficacy for the serological diagnosis of HIV-2, which will allow considering this peptide for diagnostic uses in systems with nitrocellulose based solid phase.

Characteristics, immunological response & treatment outcomes of HIV-2 compared with HIV-1 & dual infections (HIV 1/ 2) in Mumbai.

Background & objectives: Information available on HIV-2 and dual infection (HIV-1/2) is limited. This study was carried out among HIV positive individuals in an urban referral clinic in Khar, Mumbai, India, to report on relative proportions of HIV-1, HIV-2 and HIV-1/2 and baseline characteristics, response to and outcomes on antiretroviral treatment (ART). Methods: Retrospective analysis of programme data (May 2006-May 2009) at Khar HIV/AIDS clinic at Mumbai, India was done. Three test algorithm was used to diagnose HIV-1 and -2 infection. Standard ART was given to infected individuals. Information was collected on standardized forms. Results: A total of 524 individuals (male=51%; median age=37 yr) were included in the analysis over a 3 year period (2006-2009) - 489 (93%) with HIV-1, 28 (6%) with HIV-2 and 7(1%) with dual HIV-1/2 infection. HIV-2 individuals were significantly older than HIV-1 individuals (P<0.001). A significantly higher proportion of HIV-2 patients and those with dual infections had CD4 counts <200 cells/µl compared to HIV-1. HIV-2 individuals were more likely to present in WHO Clinical Stage 4. Of the 443 patients who were started on ART, 358 (81%) were still alive and on ART, 38 (8.5%) died and 3 were transferred out. CD4 count recovery at 6 and 12 months was satisfactory for HIV-1 and HIV-2 patients on protease inhibitor based regimens while this was significantly lower in HIV-2 individuals receiving 3 nucleoside reverse transcriptase inhibitors. Interpretation & conclusions: In an urban HIV clinic in Mumbai, India, HIV-2 and dual infections are not uncommon. Adaptation of the current national diagnostic and management protocols to include discriminatory testing for HIV types and providing access to appropriate and effective ART regimens will prevent the development of viral resistance and preserve future therapeutic options.

Vertical Transmission of HIV-An Update.

One of the greatest successes in AIDS research to date has by far been the discovery of successful interventions that interrupt the transmission of HIV from mother to child. It is however important to note that these successes have occurred largely in countries with great resources and the least burden of perinatal transmission of HIV. In the developing world wherein currently 95% of vertical transmission of HIV occurs, it is highly condemnable that still every minute an infected infant is said to be born in spite of the fact that vertical transmission is largely preventable, mainly because translating knowledge into practice is not always possible or feasible; This has led to a continuous growing numbers of children with HIV, thereby making pediatric HIV a looming problem rapidly draining the already burdened health care system of these countries. It is the need of the hour to appropriately address the challenges to achieve zero percent transmission of HIV from an infected mother to her child thereby giving a hope for an AIDS-free new generation worldwide.

Frequency of HIV type 2 infections among blood donor population from India: A 10-year experience.

Purpose: In India, HIV-2 epidemic is alongside with HIV-1. Blood banks are introducing nucleic acid testing (NAT) for screening. The limitation of NAT systems is the inability to detect HIV-2. Materials and Method : An analysis of HIV screening of a blood bank at a tertiary care center from 1998 to 2007 was carried out. Results : A total of 175026 donors were screened by serological assays and 789 were reactive for HIV antibody. Only 478 (61%) were confirmed positive by Western blot/immunoblot. There were 465 (97.2%) donations positive for HIV-1, 6 (1.3%) for HIV-2 (monotypic infection) and 7 (1.5%) for HIV-1 and HIV-2 (dual infection). Conclusion : We show the presence of HIV-2 infection among the blood donors and the need for incorporating HIV-2 detection also in the NAT systems.

Construction of HIV-2 gag recombinant fowlpox virus and its immune reactivity in mice / 中华传染病杂志

Objective To provide experimental evidences for developing a safe and effective re- combinant fowlpox virus which can prevent the infection of HIV-2.Methods A fowlpox virus(FPV) transferring vector was constructed by inserting HIV-2 gag gene to the downstream of a synthetic complex promoter ATI-p7.5?20 of vector pUTA2.Transfection was then carried out,and recombi- nant FPV(rFPV)was screened by 5'-bromo-deoxyuridine(BrdU),genome PCR and western blot detection.Balb/c mice were immunized with rFPV by muscular injection.Anti-HIV-2 antibody, CD4~+ and CD8~+ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA,FACS and LDH release assay,respectively.Results A transferring vector pA- gag was constructed and confirmed by amplifying a fragment of 766 bp from the rFPV genome.Mean- while,HIV-2 multi-antibody-specific protein blot(55 000)was detected from the recombinant virus and the HIV-2 specific antibody was detected from the immunized Balb/c mice.HIV-2 specific target- killing activity of spleen CTL was observed in immunized mice.Conclusion A recombinant fowlpox virus expressing HIV-2 structural protein Gag has been obtained,and it can stimulate HIV-2-specific eelluar and humoral immune reactions in mice.