HIV P24 antigen among HIV antibody seronegative blood donors in Osogbo Osun State, South Western Nigeria Olugbenga Adekunle Olowe1, Victor Olatunji Mabayoje2, Olusola Akanbi1, Olusolabomi Jose Adefioye1, Rita Ayanbolade Olowe1, Emmanuel Kehinde Fadeni3, Adeolu Sunday Oluremi3, Oluyinka Oladele Opaleye1 Department of Medical Microbiology and Parasitology, College of Health Sciences, Osogbo, Nigeria, Department of Hematology and Blood Transfusion, College of Health Sciences, Osogbo, Nigeria, 3Department of Biomedical Laboratory Sciences, Mercyland Wing, College of Health Sciences, Osogbo, Nigeria
1
2
Background: Efforts to curb the spread of HIV transmission through transfusion of blood and its products is still a problem because of challenge in countries using antibody-based rapid methods to detect infection during window period. Transmission of HIV through infected blood and its products accounts for approximately 10% in African region. Methods: This study analyzed true negativity of HIV infection in blood donors screened by ELISA test based on p24 core antigen detection. Four hundred and eighty (480) blood donors initially negative for HIV antibody by rapid screening kit, Determine™ HIV-1/2 (Abbott Laboratory, IL, USA) and re-screened with Immuno Comb® II HIV 1 and 2 (Bispot kit PBS Organics and Israel 2005). The samples were further tested for the presence of HIV antibody and p24 HIV core antigen using ELISA kits (Genscreen TM ULTRA HIV Ag–Ab) following manufacturer’s instructions. All donors initially tested negative for Hepatitis B virus, Hepatitis C virus. Result: Two (0.42%) of 480 blood donors tested positive for the p24 HIV core antigen. The two positive donors for the p24 antigen had multiple sexual partners and recent sexually transmitted infections. Conclusion: The association of the HIV p24 antigen with blood donation was highly significant (p = 0.000) and pose a great risk to recipients if screening of blood donor is only carried out by HIV antibody detection. Keywords: HIV P24 antigen, HIV Antibody, Seronegative blood donors, Immuno Comb® II HIV 1 and 2, ELISA
Background
The major routes of transmission of HIV involves sexual contact, transfusion of blood and its products while the epidemiological study of WHO in year 2000 shows those at risk of infection are homosexual or bisexual males, intravenous drug abusers, sexual contacts of infected individuals and infants of infected mother1 The Prevalence of HIV antibody in Osogbo, Nigeria is 3.2%.2 In University College Hospital, Ibadan, Nigeria, transmission of HIV through infected blood and its products accounts for approximately 62.0%.3 Blood safety remains an issue of major concern in transfusion medicine in developing countries like Nigeria where national blood transfusion services, appropriate infrastructure, trained personnel and financial resources are inadequate due to poor budgetary allocation to the health sector. Sensitive tests selection will increase blood safety decreasing the window period and this can be achieved with the use of third generation Correspondence to: Oluyinka Oladele Opaleye. Department of Medical Microbiology and Parasitology, College of Health Sciences, P.M.B. 4400, Osogbo, Osun State. Nigeria. Email: [email protected]
© 2016 Informa UK Limited, trading as Taylor & Francis Group DOI 10.1080/20477724.2016.1205311
ELISA tests, this will decrease window period by 3 weeks from the previously reported period of 6–8 weeks.4 To detect HIV infection earlier, other tests such as the antigen capture test and the polymerase chain reaction test should be done. The US Food and Drug Administration recommended in August 1995 that all donated blood and its products should be screened for HIV-1 p24 antigen, effective within 3 months of licensure of a test labeled for such use. This is expected to reduce the window period by 6 days and thus reduce the number of otherwise undetected infectious donations by approximately 25% per year.5 Testing of blood donors is carried out majorly using antibody detection kits. These kits detect antibodies to HIV antigens which appear usually later than the p24 antigen. P24 is an important structural component of the retroviral particle and estimated to be present at 2,000– 4,000 molecules in each virion.6 P24 antigen testing is sensitive and specific in diagnosing pediatric HIV infection, infection in the window phase, prediction of CD4+ T cell decline and clinical progression at early and late stage of infection, and suitable for antiretroviral treatment
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monitoring in both adults and children. Notably, p24 antigen was measurable even in patients with stably suppressed viremia and its concentrations were correlated negatively with the concentrations of CD4+ T cells and positively with the concentrations of activated CD8+ T cell subsets.7 Blood screened HIV negative by the HIV antibody detection methods alone are not completely certified free of HIV infection.8 In the past decade, combo assays have replaced stand alone p24 antigen screening and therefore more cost effective and deserve re-evaluation in the Nigerian context. This study was therefore carried out to analyze the frequency of HIV infection in their antigenemic window period among blood donors using the presence or absence of p24 core antigen in blood donors already screened as HIV negative by the antibody detection method.
Methods
Four hundred and eighty blood donors who tested negative to HIV-antibody, HBsAg and Hepatitis C virus (HCV)antibody were recruited after informed consent questionnaire and counseling for this study. HIV status of donors were determined by immunochromatographic Determine test kit with 97.96% specificity and 100% sensitivity (HIV1/2) (ABBOTT-laboratory, IL, USA), and later re-screened with Immuno Comb® II HIV 1 and 2 (Bispot kit PBS Organics and Israel 2005) with 99.70% specificity and 100% sensitivity. HBsAg status was determined with an immunochromatographic third generation Clinotech HBsAg test strips (Clinotech Diagnostics, Canada) with a sensitivity of 99.8% and specificity of 100%. Anti-HCV was similarly tested using anti-HCV strip (Clinotech Diagnostics, Canada).
Table 1 Age distribution with sex, education, occupation and number of sex partners Age group 11–20 21 (4.4%) 21–30 204 (42.5) 31–40 182 (37.9%) 41–50 61 (12.7%) 51–60 12 (2.5%) Educational level Primary 6 (1.3%) Secondary 97 (20.2%) Tertiary 377 (78.5%) Occupation Civil servant 137 (28.6%) Self employed 267 (55.6%) Unemployed 76 (15.8%) No of Sex partner One 398 (82.9%) Multiple 82 (17.1%) Age distribution with Ethnicity status of the donors 11–20 21–30 31–40 41–50 51–60
21 204 182 61 12
Male 396 (82.5%)
Female 84 (17.5%)
18 (85.7%) 169 (82.8%) 154 (84.6%) 45 (73.8%) 10 (83.3%)
3 (14.3%) 35 (17.2%) 28 (15.4%) 16 (26.2%) 2 (16.7%)
Yoruba 21 (100%) 189 (92.7%) 177 (97.3%) 61 (100%) 12 (100%) 460 (95.8%)
Non-Yoruba 0 15 (7.3%) 5 (2.7%) 0 0 20 (4.2%)
Christianity 10 (47.6%) 131 (64.2%) 112 (61.5%) 31 (50.8%) 9 (75%)
Islam 11 (52.4%) 73 (35.8%) 70 (38.5%) 30 (49.2%) 3 (25%)
Single 21 (100%) 168 (82.4%) 3 (1.6%) 0 0
Married 0 36 (17.6%) 179 (98.4%)
P-24 antigen positive 0 1(0.5%) 0 0 1(8.3%)
P-24 antigen negative 21(100%) 203 182(100%) 61(100%) 11(91.7%)
Age distribution with Religion status of the donors 11–20 21 21–30 204 31–40 182 41–50 61 51–60 12 Age distribution with sex with marital status of the donors 11–20 21 21–30 204 31–40 182 41–50 61 51–60 12 Age distribution with the donors’ p24 antigen status 11–20 21–30 31–40 41–50 51–60
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21 204 182 61 12
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Adekunle et al. HIV P24 antigen among HIV antibody seronegative blood donors
The donors were enrolled at Ladoke Akintola University Teaching Hospital, Osogbo, Osun State, Nigeria. A structured questionnaire was developed and administered to have the demographical, blood transfusion history, risky behaviors, sexual partners, drug injection history and clinical background of the donors. The study was approved by the Ethical Committee of the Ladoke Akintola University Teaching Hospital. Informed written consent was obtained from all participants of the study. About 5 ml of blood was collected from each participant into EDTA bottles, the plasma was separated and the specimens were stored at −20 °C and the screening was performed within 7 days. The samples were screened for the presence of p24 viral antigen using an ELISA kit (Genscreen™ ULTRA HIV Ag– Ab). It is a cohort study carried out over a period of four months and the statistical analysis was computed using SPSS 20 package. The results of this study were analyzed using cross tabulations to explore statistical relationship between variables and Chi square test to explore proportional relationship between groups. The level of statistical significance was set at p
1
2
Background: Efforts to curb the spread of HIV transmission through transfusion of blood and its products is still a problem because of challenge in countries using antibody-based rapid methods to detect infection during window period. Transmission of HIV through infected blood and its products accounts for approximately 10% in African region. Methods: This study analyzed true negativity of HIV infection in blood donors screened by ELISA test based on p24 core antigen detection. Four hundred and eighty (480) blood donors initially negative for HIV antibody by rapid screening kit, Determine™ HIV-1/2 (Abbott Laboratory, IL, USA) and re-screened with Immuno Comb® II HIV 1 and 2 (Bispot kit PBS Organics and Israel 2005). The samples were further tested for the presence of HIV antibody and p24 HIV core antigen using ELISA kits (Genscreen TM ULTRA HIV Ag–Ab) following manufacturer’s instructions. All donors initially tested negative for Hepatitis B virus, Hepatitis C virus. Result: Two (0.42%) of 480 blood donors tested positive for the p24 HIV core antigen. The two positive donors for the p24 antigen had multiple sexual partners and recent sexually transmitted infections. Conclusion: The association of the HIV p24 antigen with blood donation was highly significant (p = 0.000) and pose a great risk to recipients if screening of blood donor is only carried out by HIV antibody detection. Keywords: HIV P24 antigen, HIV Antibody, Seronegative blood donors, Immuno Comb® II HIV 1 and 2, ELISA
Background
The major routes of transmission of HIV involves sexual contact, transfusion of blood and its products while the epidemiological study of WHO in year 2000 shows those at risk of infection are homosexual or bisexual males, intravenous drug abusers, sexual contacts of infected individuals and infants of infected mother1 The Prevalence of HIV antibody in Osogbo, Nigeria is 3.2%.2 In University College Hospital, Ibadan, Nigeria, transmission of HIV through infected blood and its products accounts for approximately 62.0%.3 Blood safety remains an issue of major concern in transfusion medicine in developing countries like Nigeria where national blood transfusion services, appropriate infrastructure, trained personnel and financial resources are inadequate due to poor budgetary allocation to the health sector. Sensitive tests selection will increase blood safety decreasing the window period and this can be achieved with the use of third generation Correspondence to: Oluyinka Oladele Opaleye. Department of Medical Microbiology and Parasitology, College of Health Sciences, P.M.B. 4400, Osogbo, Osun State. Nigeria. Email: [email protected]
© 2016 Informa UK Limited, trading as Taylor & Francis Group DOI 10.1080/20477724.2016.1205311
ELISA tests, this will decrease window period by 3 weeks from the previously reported period of 6–8 weeks.4 To detect HIV infection earlier, other tests such as the antigen capture test and the polymerase chain reaction test should be done. The US Food and Drug Administration recommended in August 1995 that all donated blood and its products should be screened for HIV-1 p24 antigen, effective within 3 months of licensure of a test labeled for such use. This is expected to reduce the window period by 6 days and thus reduce the number of otherwise undetected infectious donations by approximately 25% per year.5 Testing of blood donors is carried out majorly using antibody detection kits. These kits detect antibodies to HIV antigens which appear usually later than the p24 antigen. P24 is an important structural component of the retroviral particle and estimated to be present at 2,000– 4,000 molecules in each virion.6 P24 antigen testing is sensitive and specific in diagnosing pediatric HIV infection, infection in the window phase, prediction of CD4+ T cell decline and clinical progression at early and late stage of infection, and suitable for antiretroviral treatment
Pathogens and Global Health 2016 VOL. 110
NOS.
4&5
205
Adekunle et al. HIV P24 antigen among HIV antibody seronegative blood donors
monitoring in both adults and children. Notably, p24 antigen was measurable even in patients with stably suppressed viremia and its concentrations were correlated negatively with the concentrations of CD4+ T cells and positively with the concentrations of activated CD8+ T cell subsets.7 Blood screened HIV negative by the HIV antibody detection methods alone are not completely certified free of HIV infection.8 In the past decade, combo assays have replaced stand alone p24 antigen screening and therefore more cost effective and deserve re-evaluation in the Nigerian context. This study was therefore carried out to analyze the frequency of HIV infection in their antigenemic window period among blood donors using the presence or absence of p24 core antigen in blood donors already screened as HIV negative by the antibody detection method.
Methods
Four hundred and eighty blood donors who tested negative to HIV-antibody, HBsAg and Hepatitis C virus (HCV)antibody were recruited after informed consent questionnaire and counseling for this study. HIV status of donors were determined by immunochromatographic Determine test kit with 97.96% specificity and 100% sensitivity (HIV1/2) (ABBOTT-laboratory, IL, USA), and later re-screened with Immuno Comb® II HIV 1 and 2 (Bispot kit PBS Organics and Israel 2005) with 99.70% specificity and 100% sensitivity. HBsAg status was determined with an immunochromatographic third generation Clinotech HBsAg test strips (Clinotech Diagnostics, Canada) with a sensitivity of 99.8% and specificity of 100%. Anti-HCV was similarly tested using anti-HCV strip (Clinotech Diagnostics, Canada).
Table 1 Age distribution with sex, education, occupation and number of sex partners Age group 11–20 21 (4.4%) 21–30 204 (42.5) 31–40 182 (37.9%) 41–50 61 (12.7%) 51–60 12 (2.5%) Educational level Primary 6 (1.3%) Secondary 97 (20.2%) Tertiary 377 (78.5%) Occupation Civil servant 137 (28.6%) Self employed 267 (55.6%) Unemployed 76 (15.8%) No of Sex partner One 398 (82.9%) Multiple 82 (17.1%) Age distribution with Ethnicity status of the donors 11–20 21–30 31–40 41–50 51–60
21 204 182 61 12
Male 396 (82.5%)
Female 84 (17.5%)
18 (85.7%) 169 (82.8%) 154 (84.6%) 45 (73.8%) 10 (83.3%)
3 (14.3%) 35 (17.2%) 28 (15.4%) 16 (26.2%) 2 (16.7%)
Yoruba 21 (100%) 189 (92.7%) 177 (97.3%) 61 (100%) 12 (100%) 460 (95.8%)
Non-Yoruba 0 15 (7.3%) 5 (2.7%) 0 0 20 (4.2%)
Christianity 10 (47.6%) 131 (64.2%) 112 (61.5%) 31 (50.8%) 9 (75%)
Islam 11 (52.4%) 73 (35.8%) 70 (38.5%) 30 (49.2%) 3 (25%)
Single 21 (100%) 168 (82.4%) 3 (1.6%) 0 0
Married 0 36 (17.6%) 179 (98.4%)
P-24 antigen positive 0 1(0.5%) 0 0 1(8.3%)
P-24 antigen negative 21(100%) 203 182(100%) 61(100%) 11(91.7%)
Age distribution with Religion status of the donors 11–20 21 21–30 204 31–40 182 41–50 61 51–60 12 Age distribution with sex with marital status of the donors 11–20 21 21–30 204 31–40 182 41–50 61 51–60 12 Age distribution with the donors’ p24 antigen status 11–20 21–30 31–40 41–50 51–60
206
21 204 182 61 12
Pathogens and Global Health 2016 VOL. 110
NOS.
4&5
Adekunle et al. HIV P24 antigen among HIV antibody seronegative blood donors
The donors were enrolled at Ladoke Akintola University Teaching Hospital, Osogbo, Osun State, Nigeria. A structured questionnaire was developed and administered to have the demographical, blood transfusion history, risky behaviors, sexual partners, drug injection history and clinical background of the donors. The study was approved by the Ethical Committee of the Ladoke Akintola University Teaching Hospital. Informed written consent was obtained from all participants of the study. About 5 ml of blood was collected from each participant into EDTA bottles, the plasma was separated and the specimens were stored at −20 °C and the screening was performed within 7 days. The samples were screened for the presence of p24 viral antigen using an ELISA kit (Genscreen™ ULTRA HIV Ag– Ab). It is a cohort study carried out over a period of four months and the statistical analysis was computed using SPSS 20 package. The results of this study were analyzed using cross tabulations to explore statistical relationship between variables and Chi square test to explore proportional relationship between groups. The level of statistical significance was set at p
