Impact of Multiplex PCR in Reducing the Risk of Residual Transfusion-Transmitted Human Immunodeficiency and Hepatitis B and C Viruses in Burkina Faso
Arzouma Paul Yooda1,2,3, Serge Theophile Soubeiga1,2, K. Yacouba Nebie3, Birama Diarra1, Salam Sawadogo3, Abdoul Karim Ouattara1,2, Dorcas Obiri-Yeboah4, Albert Theophane Yonli1,2, Issoufou Tao1,2, Pegdwende Abel Sorgho1,2, Honorine Dahourou3 and Jacques Simpore1,2.
of Molecular Biology and Molecular Genetics (LABIOGENE) UFR/SVT,
University Ouaga I Prof. Joseph KI-ZERBO, 03 BP 7021 Ouagadougou
03, Burkina Faso.
2 Biomolecular Research Center Pietro Annigoni (CERBA), 01 BP 364 Ouagadougou 01, Burkina Faso.
3 National Blood Transfusion Center (CNTS), 01 BP 5372 Ouagadougou 01, Burkina Faso.
4 Department of Microbiology and Immunology, School of Medical Sciences, University of Cape Coast, Ghana
Received: April 13, 2018
Accepted: June 12, 2018
Mediterr J Hematol Infect Dis 2018, 10(1): e2018041 DOI 10.4084/MJHID.2018.041
This article is available on PDF format at:
| This is an Open Access article distributed
under the terms of the Creative Commons Attribution License
(https://creativecommons.org/licenses/by-nc/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Background and Objective:
The improved performance of serological tests has significantly reduced
the risk of human immunodeficiency and hepatitis B and C viruses
transmission by blood transfusion, but there is a persistence of
residual risk. The objective of this study was to evaluate the impact
of multiplex PCR in reducing the risk of residual transmission of these
viruses in seronegative blood donors in Burkina Faso.
Our study showed a high residual risk of HBV transmission through blood
transfusion. Due to the high prevalence of blood-borne infections in
Burkina Faso, we recommend the addition of multiplex PCR to serologic
tests for optimal blood donation screening.
Today, the risk of transmission of these viruses through transfusion has markedly decreased due to the application of preventive measures and improved performance of serological tests. Nevertheless, there is still a persistence of a residual risk mainly related to the serological window periods characterized by low-level serological viral markers, which are usually undetectable by conventional serological assays. In order to ensure blood safety, the World Health Organization (WHO) recommends the recruitment of voluntary, regular and unpaid donors selected from low-risk populations.
In Burkina Faso, since its operationalization in 2005, Regional Blood Transfusion Center of Ouagadougou (CRTS/O) has continuously improved its blood transfusion safety policy by applying measures aimed at reducing the residual risk of transfusion, namely: medical selected donors, recruitment of voluntary, regular and unpaid donors and the use of fourth-generation serological tests for the biological qualification of blood donations.
With a view to optimal transfusion safety research, Burkina Faso is now considering the introduction of PCR in the screening of blood donations. The application of this technology in blood transfusion in developed countries has made it possible to reduce the residual transfusion risk associated with the serological window.[12-14] Burkina Faso, a developing country (DC) has a high endemicity of these transfusion-transmitted viral infections but has very little data on the impact of multiplex PCR in our context. The present study was conducted at the CRTS/O to evaluate the impact of multiplex PCR in reducing the risk of residual transfusion-transmitted HIV,
HBV and HCV infections in blood donors seronegative for these three viruses in Burkina Faso.
Material and Methods
Ethical considerations. Ethical approval for the study was obtained from Ethics Committee for Health Research of Burkina Faso (deliberation n° 2015-6-080). Written informed consent was provided by all study participants.
Sampling. Venous blood specimens were collected from blood donors on a red-top and EDTA tubes. After centrifugation at 1500 g for 20 minutes, aliquots of serum and plasma were performed within 6 hours after samples collection for laboratory analyzes.
Screening for HIV, HBV and HCV serological markers. The serological markers of HIV, HBV and HCV were investigated on donor sera by chemiluminescent microparticle immunoassay (CMIA) using ARCHITECT i1000 SR (Abbott Diagnosis, USA). The p24 antigen and antibodies against HIV1/2 were simultaneously investigated using the ARCHITECT HIV Ag/Ab Combo Kit (Abbott, Wiesbaden Germany). HBV surface antigen (HBsAg) and anti-HCV antibodies were respectively determined using the ARCHITECT HBsAg Qualitative II (Abbott, Ireland, Sligo Ireland) and ARCHITECT Anti-HCV (Abbott GmbH & Co.KG, Wiesbaden Germany).
Detection for viral genomes of HIV, HBV and HCV. The detection of viral nucleic acid was performed sequentially on the samples obtained exclusively from blood donors who tested seronegative for the three viruses (HIV, HBV, HCV). A first multiplex PCR was carried out on mini-pools of 8 plasma samples each. The mini- pools detected positive were reconstituted into 2 mini-pools of 4 plasma samples for a second multiplex PCR. Finally, an individual PCR assay was performed on the samples from positive mini- pools to the second PCR.
Constitution of mini-pools. Seronegative tested samples were divided into mini-pools of 8 plasma samples each. Each mini-pool consisted of 160 μL of plasma from each donation (i.e. 1.28 mL of plasma per mini-pool). Mini-pools of 8 plasma tested positive at the first assay were reconstituted into mini-pools of 4 plasma samples of 100 μL of each donation (i.e. 400 μL per mini-pool).
Extraction and amplification by multiplex PCR in real time. Molecular analysis was performed at the laboratory of Molecular Biology and Genetics (CERBA/ LABIOGENE). Extractions of viral nucleic acid were performed with 100 μL of mini- pool plasma using the Ribo-sorb sacaceTM Kit (Sacace Biotechnologies®, Como Italy). PCR amplification was performed on the SaCycler-96 Real Time PCR v.7.3 (Sacace Biotechnologies) with the HCV/HBV/HIV Real-TM sacaceTM multiplex kit (Sacace Biotechnologies®, Como, Italy). The PCR was carried out using a reaction volume of 25 μL (10 μL of DNA/RNA and 15 μL of the mix). The SaCycler-96 real-time multiplex PCR amplification was performed according to the following program: 1 cycle of 95°C for 15 s follows by 46 cycles of 95 °C for 15 s and 60°C for 40 s. The sensitivity of the HCV/HBV/HIV Real-TM kit was respectively 10 IU/mL, 5 IU/mL and 20 copies/mL for HCV, HBV and HIV. After the first two amplifications, individual real-time PCR for HBV-positive samples of the mini-pools was performed to find the positive sample (s) from each pool. For this step the sacaceTM Ribo-Sorb Silica kit was used for extraction and the sacaceTM HBV Real-TM Qual kit was used for amplification according to the manufacturer's recommendations.
Statistical analyzes. The data were analyzed using the Epi Info version 7 software. The chi-square test was used for comparisons and any value was considered statistically significant for p ≤ 0.05.
|Table 1. Seroprevalences of HIV, HBV and HCV according to socio-demographic characteristics of blood donors at CRTS / O.|
Of 989 donations tested for antibodies and/or antigens associated with HIV, HBV and HCV infection, 865 (87.46%) donations were detected seronegative versus 124 (12.54%) seropositive cases. HIV infection was positive in 2.53% (25/989) of blood donors, HBV in 7.28% (72/989) and HCV in 2.73% (27/989) of cases. HIV/HBV, HBV/HCV, HIV/HCV and HIV/HBV/HCV
coinfections were 0.4% (4/989), 0.71% (7/989), 0.2% (2/989) and 0.1% (1/989) respectively. The seroprevalence of transfusion-transmissible infections was higher among first-time donors and mobile collection sites (Table 2).
|Table 2. Distribution of HBV DNA according to the socio-demographic characteristics of the 38 donors screened positive for multiplex PCR.|
Prevalence of HIV, HBV, and HCV genomes (DNA, RNA) in seronegative screened blood donors by the CMIA method. Forty-three (43) mini-pools were detected positive for HBV out of the 101 mini-pools of 8 plasma samples tested at the first real-time multiplex PCR. No mini-pool was positive for HIV and HCV. Of 86 mini-pools of 4 plasma samples reconstituted for the second multiplex PCR, 42 were positive for HBV. No mini-pool was positive for HIV and HCV. Of the 168 (42 x 4) samples tested by single PCR, 38 were positive for HBV. Altogether, 4.70% (38/808) of seronegative donations were positive by multiplex PCR. These positive samples were confirmed using the same detection kit on the same device.
The donations came exclusively from voluntary and unpaid blood donors. In recent years, family or replacement donations have been phased out by the CRTS/O in order to comply with WHO recommendations for better blood safety. Nevertheless, our study reported seroprevalences of 2.53%, 7.28%, and 2.73% respectively for HIV, HBV, and HCV. The seroprevalence of 2.53% of HIV reported in this study is similar to that of 2.00% reported in the general population of the central region of Burkina Faso in 2016 where our study was conducted and it is markedly higher than the 0.8% reported in the general population of Burkina Faso by the same study. It is also higher than the 1.8% reported in blood donors from three regional centers in Burkina Faso in 2009 but comparable to the 2.21% reported in Koudougou blood donors in 2012. As for HBV, the prevalence of 7.28% obtained in our study was lower than those found in previous studies in the general population9 and blood donors in Nouna, Ouagadougou and Koudougou or in specific groups (pregnant women and health workers). The prevalence of 2.73% of HCV infection was also lower than the 4.4% prevalence reported among blood donors in 2014. Altogether, the results of this study show a high prevalence of HIV and HCV against a low prevalence of HBV compared to previous studies in Burkina Faso. This could be explained by the low rate (only 11.3%) of regular blood donor in Burkina Faso.
This decrease in HBV prevalence can be attributed to extensive awareness and screening campaigns during the last decade, significant improvement in the accessibility and availability of hepatitis B vaccine and expanded immunization program against HBV for children at 8 weeks after birth since 2006. In addition, awareness campaigns, which are much more focused on hepatitis B infection with free screening in recent years, are increasingly providing a selective population of voluntary donors at fixed sites who already know their negative serological status for hepatitis B infection but generally unaware of their serology for HIV and/or HCV. This could also explain the high seroprevalence of the latter viruses and low prevalence of HBV infection compared to previous data in Burkina Faso. Furthermore, unpublished data have shown a variation in the prevalence of HBV infection between different districts of Ouagadougou. This observation could be extended to HIV and/or HCV infections. In this study, the seroprevalences of these three viruses were higher in first-time donors than in repeat donors. A similar observation has been made in several studies carried out in Burkina Faso and other West African countries.[16,17] The high prevalence of these viral infections in first-time donors can be attributed to the lower level of knowledge about blood-borne infections and routes of transmission compared to regular or repeat donors. However, to cope with the high demand for labile blood products (LBP), mobile blood collections by CRTS/O is required in high schools, universities, barracks, places of worship etc. These mobile collections would encourage the enrollment of first-time donors although the latter, unlike regular donors, are not always aware of the issue of transfusion safety. This increases the risk of collection of infected donors during seroconversion period. In the present study, 63.90% of blood donors come from mobile sites explaining the predominance of first- time donors (55.21%).
Likewise, the seroprevalences of these three infections were higher in the blood donors at the mobile sites compared to those taken at the fixed site at the CRTS/O with a statistically significant difference (p < 0.001) for all these three viral infections. This observation suggests that our method of medical donor selection is not adapted to our blood donors’ recruitment policy in mobile sites. Indeed, in mobile sites, blood collections are often organized without prior awareness-raising of potential donors on transfusion safety issues. This is especially true as mobile collections tend to encourage the recruitment of first-time donors. All of these results show that, despite the medical selection of blood donors and the computerization of their data, seroprevalences of HIV, HBV and HCV remain very high among blood donors in Burkina Faso. This reflects shortcomings in the promotion of blood donation and donor education prior to blood collection hence the low proportion of regular donors. These high seroprevalences of HIV, HBV and HCV in a population of volunteer donors also pose the problem of the quality of medical selection. Indeed, a well-conducted medical selection with relevant selection criteria is highly effective in controlling the risk associated with the silent period in infections.[19-21] In Burkina Faso, the pre-donation questionnaire and the quality of the selection have not yet been formally evaluated. Nevertheless, a study by Kafando et al. showed that there was no significant difference between the positivity rate among donors accepted to donate and those who were refused.
Of 808 seronegative HIV, HBV, and HCV donations tested by multiplex PCR, 38 (4.70%) residual cases were detected positive. These results are higher than those obtained in similar studies in other developing countries also experiencing high prevalence of these three infections. For example: in Ghana, out of 9372 seronegative screened donors by rapid tests, 3% were detected HBV-positive DNA; no cases of HIV and HCV were detected.
In South Africa, the residual risk rate of HIV, HBV and HCV was estimated to be 1 per 45,765, 1 per 11, 810 and 1 per 732, 200; in Thailand, out of 4798 HIV-negative donations, 6 cases of HBV- positive cases were detected, no HCV and HIV cases were detected. Our results are considerably higher compared to those obtained in developed countries where the prevalence of these three infections is low. For example: United States (HIV: 1 / 2million, HCV: 1 / 270,000), Italy (HCV: 2.5 / million, HIV: 1.8 / million, HBV: 57.8 / million), Germany (HCV: 1 / 10.8 million, HBV: 1/360 000). All 38 donations tested positive by multiplex PCR were only positive for HBV DNA.
These results suggest that the residual risk of transmission of HBV by blood transfusion is greater compared to that of HIV and HCV in Burkina Faso. Otherwise, the contribution of PCR would be more important for HBV compared to HIV and HCV in our country. The proportion of positive HBV donations that are only detectable in the PCR (4.72%) obtained in our study can be explained by the high prevalence and incidence of this infection in the blood donor population in Burkina Faso. These high prevalences and incidences are necessarily accompanied by significant proportions of infections in the early phase from the point of view of serological markers. Added to this is the high prevalence of occult hepatitis B reported in several studies in Burkina Faso.[28,29] Occult hepatitis B is characterized by the presence of HBV DNA in the serum of a patient who is screened for HBsAg by the usual serological tests. Although our study did not allow us to detect residual cases associated with HIV and HCV, the number of donations detected positive HBV by PCR confirms that PCR is more sensitive than ELISA. This sensitivity had already been demonstrated by some studies conducted in blood donors in Burkina Faso, and in two neighboring countries, Togo and Ghana. The absence of positive cases of HIV and HCV in our study could be explained by the relatively low prevalence of these viruses in the general population but also the limited size of our sample. The residual risk of HIV estimated by a mathematical model based on serological tests was 1 for 55,000 donations among blood donors in Burkina Faso in 2011. Similarly, no cases of residual HIV and HCV risk detectable by PCR were reported in two other similar blood donor studies in Ghana, a neighboring country in Burkina Faso and Kenya, a country of Sub-Saharan Africa like ours. However, cases of residual risk of HIV and HCV, although relatively low compared to HBV, have been observed in similar large-sample studies in other developing countries and developed countries.[13,26,27]
Of the 38 cases of HBV DNA detected in our study, 24 (63.2%) were first-time donors versus 14 (36.8%) who were repeat donors. These results show that the residual risk of transfusion is higher in first-time donors compared to repeat donors, but without a significant statistical difference. These results confirm are consistent with the literature that donations from regular donors would be the most safety. This is a cornerstone of WHO's strategy of promoting regular donations. Nevertheless, we note in our study that residual cases of HBV are also important in repeat donors. From a mathematical model, Nagalo et al. estimated equally high incidence rates of 3270.2, 5874.1, and 6784.6 per 100,000 donations for HIV-1, HBV, and HCV, respectively, among repeat donors. Considering a 100% HBV transmission rate by transfusion of a contaminated LBP, the PCR prevented 38 cases of HBV transmission or even more if these 38 donations were used for the preparation of other LBP such as Frozen Fresh Plasmas (FFPs) and Standard Platelet Concentrates (SPC). All of these results show that in addition to promoting unpaid and regular voluntary donation, the implementation of the PCR is useful in the context of Burkina Faso.
- UNAIDS. Global Report on AIDS Epidemic: Treatment and care. 2008. Available at www.unaids.org/sites/default/files/media_asset/jc1510_2008globalrep ort_en_0.pdf. Accessed on 09/04/2018.
S, Chalabi Z, Perel P, Guerriero C, Roberts I. The risk of
transfusion‐transmitted infections in sub‐Saharan Africa. Transfusion.
2010;50:433-42. https://doi.org/10.1111/j.1537-2995.2009.002402.x PMid:19843290
- Fasola F, Otegbayo I. Post-transfusion viral hepatitis in sickle cell anaemia: retrospective-prospective analysis. Nigerian Journal of Clinical Practice. 2002;5:16-9.
- Nagalo BM, Bisseye C, Sanou M, Kienou K, Nebié YK, Kiba A, Dahourou H, Ouattara S, Nikiema JB, Moret R. Seroprevalence and incidence of transfusion‐transmitted infectious diseases among blood donors from regional blood transfusion centres in Burkina Faso, West Africa. Tropical medicine & international health. 2012;17:247- 53. https://doi.org/10.1111/j.1365-3156.2011.02902.x PMid:21988100
- Nagalo MB, Sanou M, Bisseye C, Kaboré MI, Nebie YK, Kienou K, Kiba A, Dahourou H, Ouattara S, Zongo JD. Seroprevalence of human immunodeficiency virus, hepatitis B and C viruses and syphilis among blood donors in Koudougou (Burkina Faso) in 2009. Blood transfusion. 2011;9:419. PMid:21839011 PMCid:PMC3200412
- Pietra V, Kiema D, Sorgho D, Kabore S, Mande S, Castelli F, Puoti M, Simpore J. Prévalence des marqueurs du virus de l'hépatite B et des anticorps contre le virus de l'hépatite C parmi le personnel du District Sanitaire de Nanoro, Burkina Faso. Science et technique, sciences de la santé. 2008;31:53-9.
- Collenberg E, Ouedraogo T, Ganamé J, Fickenscher H, Kynast‐Wolf G, Becher H, Kouyaté B, Kräusslich HG, Sangaré L, Tebit DM. Seroprevalence of six different viruses among pregnant women and blood donors in rural and urban Burkina Faso: a comparative analysis. Journal of medical virology. 2006;78:683-92. https://doi.org/10.1002/jmv.20593 PMid:16555290
- Zeba MTA, Sanou M, Bisseye C, Kiba A, Nagalo BM, Djigma FW, Compaoré TR, Nebié YK, Kienou K, Sagna T. Characterisation of hepatitis C virus genotype among blood donors at the regional blood transfusion centre of Ouagadougou, Burkina Faso. Blood Transfusion. 2014;12:s54. PMid:24599906 PMCid:PMC3934227
- Tao I, Compaoré TR, Diarra B, Djigma F, Zohoncon TM, Assih M, Ouermi D, Pietra V, Karou SD, Simpore J. Seroepidemiology of hepatitis B and C viruses in the general population of burkina faso. Hepatitis research and treatment. 2014;2014.
- Kleinman S, Busch MP, Korelitz JJ, Schreiber GB. The incidence/window period model and its use to assess the risk of transfusion-transmitted human immunodeficiency virus and hepatitis C virus infection. Transfusion medicine reviews. 1997;11:155-72. https://doi.org/10.1053/tmrv.1997.0110155 PMid:9243769
- WHO. Blood safety: A strategy for the AFRICAN REGION. 2001. Available at http://apps.who.int/iris/bitstream/handle/10665/95734/AFR_RC51_R2.pdf?sequence=1&isAllowed=y. Accessed on 09/04/2018.
- Davidson T, Ekermo B, Gaines H, Lesko B, Åkerlind B. The cost‐ effectiveness of introducing nucleic acid testing to test for hepatitis B, hepatitis C, and human immunodeficiency virus among blood donors in Sweden. Transfusion. 2011;51:421-9. https://doi.org/10.1111/j.1537-2995.2010.02877.x PMid:20849409
- Stramer S, Glynn S, Kleinman S. Detection of Hiv-1 and Hcv infections among antibody-negative blood donors by nucleic acid amplification testing. Vox Sanguinis. 2005;88:68-9.
- Zou S, Dorsey KA, Notari EP, Foster GA, Krysztof DE, Musavi F, Dodd RY, Stramer SL. Prevalence, incidence, and residual risk of human immunodeficiency virus and hepatitis C virus infections among United States blood donors since the introduction of nucleic acid testing. Transfusion. 2010;50:1495-504. https://doi.org/10.1111/j.1537-2995.2010.02622.x PMid:20345570
- CNLS/IST. Rapport d'activité sur la riposte au SIDA au Burkina Faso. http://www.unaidsorg/sites/default/files/country//BFA_narrative_repo rt_2016pdf. 2016. Accessed on 09/04/2018.
- Mavenyengwa RT, Mukesi M, Chipare I, Shoombe E. Prevalence of human immunodeficiency virus, syphilis, hepatitis B and C in blood donations in Namibia. BMC Public Health. 2014;14:424. https://doi.org/10.1186/1471-2458-14-424 PMid:24884633 PMCid:PMC4012713
- Noubiap JJN, Joko WYA, Nansseu JRN, Tene UG, Siaka C. Sero- epidemiology of human immunodeficiency virus, hepatitis B and C viruses, and syphilis infections among first-time blood donors in Edéa, Cameroon. International Journal of Infectious Diseases. 2013;17:e832-e7. https://doi.org/10.1016/j.ijid.2012.12.007 PMid:23317526
- Nébié K, Olinger C, Kafando E, Dahourou H, Diallo S, Kientega Y, Domo Y, Kienou K, Ouattara S, Sawadogo I. Faible niveau de connaissances des donneurs de sang au Burkina Faso; une entrave potentielle à la sécurité transfusionnelle. Transfusion clinique et biologique. 2007;14:446-52. https://doi.org/10.1016/j.tracli.2007.12.005 PMid:18295528
- Stokx J, Gillet P, De Weggheleire A, Casas EC, Maendaenda R, Beulane AJ, Jani IV, Kidane S, Mosse CD, Jacobs J. Seroprevalence of transfusion-transmissible infections and evaluation of the pre- donation screening performance at the Provincial Hospital of Tete, Mozambique. BMC infectious diseases. 2011;11:141. https://doi.org/10.1186/1471-2334-11-141 PMid:21605363 PMCid:PMC3120673
- Seck M, Dieye B, Guèye Y, Faye B, Senghor A, Toure S, Dieng N, Sall A, Touré A, Dièye T. Évaluation de l'efficacité de la sélection médicale des donneurs de sang dans la prévention des agents infectieux. Transfusion Clinique et Biologique. 2016;23:98-102. https://doi.org/10.1016/j.tracli.2015.11.001 PMid:26681660
- Tagny CT, Kouao MD, Touré H, Gargouri J, Fazul AS, Ouattara S, Anani L, Othmani H, Feteke L, Dahourou H. Transfusion safety in francophone African countries: an analysis of strategies for the medical selection of blood donors. Transfusion. 2012;52:134-43. https://doi.org/10.1111/j.1537-2995.2011.03391.x PMid:22014098 PMCid:PMC3668689
- Kafando E, Nébié Y, S S, Kienou K, Dahourou H, Simpore J. Risk Behavior among Ineligible Blood Donors in a Blood Transfusion Center (Burkina Faso). J Hematol Blood Transfus Disord. 2017;4:015.
- Owusu‐Ofori S, Temple J, Sarkodie F, Anokwa M, Candotti D, Allain JP. Predonation screening of blood donors with rapid tests: implementation and efficacy of a novel approach to blood safety in resource‐poor settings. Transfusion. 2005;45:133-40. https://doi.org/10.1111/j.1537-2995.2004.04279.x PMid:15660820
- Vermeulen M, Lelie N, Sykes W, Crookes R, Swanevelder J, Gaggia L, Le Roux M, Kuun E, Gulube S, Reddy R. Impact of individual‐ donation nucleic acid testing on risk of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus transmission by blood transfusion in South Africa. Transfusion. 2009;49:1115-25. https://doi.org/10.1111/j.1537-2995.2009.02110.x PMid:19309474
- Nantachit N, Thaikruea L, Thongsawat S, Leetrakool N, Fongsatikul L, Sompan P, Fong YL, Nichols D, Ziermann R, Ness P. Evaluation of a multiplex human immunodeficiency virus‐1, hepatitis C virus, and hepatitis B virus nucleic acid testing assay to detect viremic blood donors in northern Thailand. Transfusion. 2007;47:1803-8. https://doi.org/10.1111/j.1537-2995.2007.01395.x PMid:17880604
- Velati C, Romanò L, Fomiatti L, Baruffi L, Zanetti AR. Impact of nucleic acid testing for hepatitis B virus, hepatitis C virus, and human immunodeficiency virus on the safety of blood supply in Italy: a 6‐ year survey. Transfusion. 2008;48:2205-13. https://doi.org/10.1111/j.1537-2995.2008.01813.x PMid:18631163
- Hourfar MK, Jork C, Schottstedt V, Weber‐Schehl M, Brixner V, Busch MP, Geusendam G, Gubbe K, Mahnhardt C, Mayr‐Wohlfart U. Experience of German Red Cross blood donor services with nucleic acid testing: results of screening more than 30 million blood donations for human immunodeficiency virus‐1, hepatitis C virus, and hepatitis B virus. Transfusion. 2008;48:1558-66. https://doi.org/10.1111/j.1537-2995.2008.01718.x PMid:18466173
- Somda K, Sermé A, Coulibaly A, Cissé K, Sawadogo A, Sombié A, Bougouma A. Hepatitis B Surface Antigen Should Not Be the Only Sought Marker to Distinguish Blood Donors towards Hepatitis B Virus Infection in High Prevalence Area. Open Journal of Gastroenterology. 2016;6:362. https://doi.org/10.4236/ojgas.2016.611039
- Diarra B, Yonli AT, Sorgho PA, Compaore TR, Ouattara AK, Zongo WA, Tao I, Traore L, Soubeiga ST, Djigma FW. Occult hepatitis B virus infection and associated genotypes among HBsAg-negative subjects in Burkina Faso. Mediterranean journal of hematology and infectious diseases. 2018;10. https://doi.org/10.4084/mjhid.2018.007
- Raimondo G, Allain J-P, Brunetto MR, Buendia M-A, Chen D-S, Colombo M, Craxì A, Donato F, Ferrari C, Gaeta GB. Statements from the Taormina expert meeting on occult hepatitis B virus infection. Journal of hepatology. 2008;49:652-7. https://doi.org/10.1016/j.jhep.2008.07.014 PMid:18715666
- Nagalo B, Bisseye C, Sanou M, Nebié Y, Kiba A, Kienou K, Zongo J, Simporé J. Diagnostic moléculaire du virus de l'immunodéficience humaine acquise (VIH) sur les pools de plasmas de donneurs de sang au centre régional de transfusion sanguine de Ouagadougou (CRST- 0), Burkina Faso. Médecine tropicale. 2011;71:137-41.
- Assih M, Feteke L, Bisseye C, Ouermi D, Djigma F, Karou SD, Simpore J. Molecular diagnosis of the human immunodeficiency, hepatitis B and C viruses among blood donors in lomé (Togo) by multiplex real time PCR. The Pan African Medical Journal. 2016;25. https://doi.org/10.11604/pamj.2016.25.242.7096
- Lefrère JJ, Dahourouh H, Dokekias AE, Kouao MD, Diarra A, Diop S, Tapko JB, Murphy EL, Laperche S, Pillonel J. Estimate of the residual risk of transfusion‐transmitted human immunodeficiency virus infection in sub‐Saharan Africa: a multinational collaborative study. Transfusion. 2011;51:486-92. https://doi.org/10.1111/j.1537- 2995.2010.02886.x PMid:20880002
- Basavaraju S, Mwangi J, Nyamongo J, Zeh C, Kimani D, Shiraishi R, Madoda R, Okonji J, Sugut W, Ongwae S. Reduced risk of transfusion‐transmitted HIV in Kenya through centrally co‐ ordinated blood centres, stringent donor selection and effective p24 antigen‐HIV antibody screening. Vox sanguinis. 2010;99:212-9. https://doi.org/10.1111/j.1423-0410.2010.01340.x PMid:20497410