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Journal of Clinical Microbiology, December 2005, p. 6218-6219, Vol. 43, No. 12
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.12.6218-6219.2005

LETTER TO THE EDITOR

Visual Detection of Multiple Viral Amplicons by Dipstick Assay: Its Application in Screening of Blood Donors a Welcome Tool for the Limited Resource Settings


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LETTER
 
Hepatitis B virus (HBV) is a significant cause of transfusion-transmitted infections. To achieve entirely HBV-safe transfusion still appears to be an unreachable objective, at least in the high-prevalence regions of the developing world. Transfusion-related HBV safety depends on the screening technique adopted by the blood banks and its prevalence in the community, and the more sensitive the screening technique adopted the less the chance of transmitting HBV infection through the transfusion (1). All endeavors towards lowering the risk of transfusion transmitted HBV and/or HCV infection play an important role in prevention of this preventable human suffering. This is particularly significant in the developing countries with limited resources and overstretched health budgets. A recently published report by Dineva et al. is a remarkable achievement in this direction (2). The authors have introduced a simple, sensitive, instrument-independent technique for visual detection of multiple nucleic acid hybridization on the dipstick membrane. The report shows high sensitivity and reproducibility of the test for detection of HBV DNA, HCV RNA, and human immunodeficiency virus RNA among blood donors by minipools. Unfortunately, the genomic screening technology is still expensive and not affordable for the developing regions of the world where these viruses are highly endemic, which not only increases the risk of transfusion-transmitted HBV and/or HCV infection but is perhaps an important mode of spread of this infection. Most of the developing countries are screening the collected units of blood for HBsAg, anti-HBc, anti-HBs, and anti-HCV by enzyme immunoassay. The blood units which are anti-HBc positive or anti-HCV positive are rejected, thus leading to high rejection rates of collected units of blood. At this center, 3.2% of units are rejected because of isolated anti-HBc positivity (4). The rejection of these invaluable collected blood units and exclusion of the munificent volunteer blood donors because of isolated anti-HBc or anti-HCV positivity lead to chronic shortage of blood. The blood units which are isolated as being anti-HBc positive may not necessarily contain HBV DNA when tested by the nucleic acid amplification testing (NAT) technology, or anti-HCV-positive units may not be positive for HCV RNA if tested by PCR. Hence, they can perhaps be utilized if the low-cost genomic screening resources are made available to the developing countries; on the other hand, the anti-HBc- and/or anti-HBs-positive donors might be positive for HBV DNA if tested by the sensitive NAT assay system (3, 5). The visual detection dipstick method for detection of HBV and HCV nucleic acid appears an attractive alternative to the intricate instrument-dependent methods for the countries with limited resources. This simple instrument-independent method is less expensive than the conventional instrument-dependent method and can be adopted by the blood banks of the developing countries where these viruses are highly endemic to possibly ensure the achievement of zero risk of transfusion-transmitted HBV and HCV infection. This will not only ensure the safe blood transfusion at lesser testing cost but will also reduce the preventable human suffering of transfusion-transmitted HBV and HCV infection.


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REFERENCES
 
    1
  1. Allain, J. -P. 2004. Occult hepatitis B virus infection: implications in transfusion. Vox Sang. 86:83-91.[CrossRef][Medline]
    2. 2
  2. Dineva, M. A., D. Chandotti, F. Fletcher-Brown, J. -P. Allain, and H. Lee. 2005. Simultaneous visual detection of multiple viral amplicons by dipstick assay. J. Clin. Microbiol. 43:4015-4021.[Abstract/Free Full Text]
    3. 3
  3. Murokawa, H., A. Yoshikawa, H. Ohnuma, A. Itawa, N. Katoh, M. Miyamoto, H. Mine, H. Emura, and K. Tadokoro. 2005. Epidemiology of blood donors in Japan, positive for hepatitis B virus and hepatitis C virus by nucleic acid amplification testing. Vox Sang. 88:10-16.[Medline]
    4. 4
  4. Panhotra, B. R., A. Bahrani, and Z. U. Hassan. 2005. Epidemiology of antibody to hepatitis B core antigen screening among blood donors in eastern Saudi Arabia: need to replace the test by HBV DNA testing. Saudi Med. J. 26:270-273.[Medline]
    5. 5
  5. Silva, C. M. D., C. Costi, and C. Costa. 2005. Low rate of occult hepatitis B virus infection among anti-HBc positive blood donors living in a low prevalence region in Brazil. J. Infect. 51:24-29.[Medline]
B. R. Panhotra*
Z. U. Hassan
C. S. Joshi
A. Bahrani
Department of Laboratory & Blood Bank
King Fahad Hospital
Al-Hofuf, Al-Hasa 31982
Saudi Arabia

* Phone: 966502910024, Fax: 96635752255, E-mail: drpanhotra{at}gmail.com


Authors' Reply


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LETTER 
 
We read with interest the letter by Panhotra et al., Department of Laboratory and Blood Bank, King Fahad Hospital, Saudi Arabia, commenting on our recent publication (4). This letter confirms that simple, robust and affordable genomic screening tools are sorely needed to improve the safety of blood supplies in resource-limited settings.

In most resource-poor countries of Africa, Asia, and Latin America, postdonation screening by enzyme immunoassay has revealed a prevalence of viral markers in blood donors ranging between 1 and 20% for anti-human immunodeficiency virus (HIV), 0.5 and 15% for anti-HCV, and 3 and 25% for HBV (1-3, 5-7). Indeed, in many developing countries, one in five apparently healthy donors is infected with one or more of these transfusion-transmitted viruses. Yet over 40% of donated blood worldwide is not screened for all relevant transfusion-transmissible infections (8). As a consequence, it has been estimated that transfusion of unsafe blood accounts for 8 to 16 million HBV infections, 2.3 to 4.7 million HCV infections, and 80,000 to 160,000 HIV infections each year. The failure to screen blood supplies for HIV, HBV, and HCV systematically is largely due to the high cost of instruments and tests and technical obstacles such as instrument repair and training of personnel.

The implementation of a visual dipstick-based triplex NAT assay will provide a means to ensure the screening of all three transfusion-transmitted viruses. In addition, this assay format can also be useful in developed countries under circumstances in which a rapid result is required. It has been found that up to 40% of the attendees of inner city clinics who have been tested for HIV do not return for their test results and are often not traceable and therefore lost to the healthcare system. A rapid and simple near-patient NAT assay could be useful in this setting by giving the physician the opportunity to make the initial intervention while the patient is still at the clinic.

In conclusion, the comment letter supports the objective of our research: making NAT available and affordable to resource-poor settings.


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REFERENCES 
 
    1
  1. Apetrei, C., I. Loussert-Ajaka, D. Descamps, F. Damond, S. Saragosti, F. Brun-Vezinet, and F. Simon. 1996. Lack of screening test sensitivity during HIV-1 non-subtype B seroconversion. AIDS 10:F57-F60.[Medline]
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  2. Basaras, M., A. Santamaria, M. Sarsa, E. Gutierez, Y. de Olano, and R. Sisterna. 1999. Seroprevalence of hepatitis B and C and human immunodeficiency type 1 viruses in a rural population from the Republic of Equatorial Guinea. Trans. R. Soc. Trop. Med. Hyg. 93:250-252.[CrossRef][Medline]
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  3. Larkin, M. 2000. WHO’s blood-safety initiative: a vain effort? Lancet 355:1245.[Medline]
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  4. Dineva, M. A., D. Chandotti, F. Fletcher-Brown, J. P. Allain, and H. Lee. 2005. Simultaneous visual detection of multiple viral amplicons by dipstick assay. J. Clin. Microbiol. 43:4015-4021.
    5. 5
  5. Mwangi, J. W. 1999. Viral markers in a blood donor population. East Afr. Med. J. 76:35-37.
    6. 6
  6. Opare-Sem, O., S. Owusu-Ofori, and J.-P. Allain. 2002. A novel approach to blood safety: pre-donation screening of blood donors for viral markers. Africa Sanguine 5:12-16.
    7. 7
  7. Sarkodie, F., M. Adarkwa, Y. Sarkodie, J. W. Acheampong, and J. P. Allain. 2001. Screening for viral markers by EIA in volunteer and replacement donors in Kumasi, Ghana. Vox Sang. 80:142-147.[CrossRef][Medline]
    8. 8
  8. World Health Organization. 2001. Global database on blood safety from 1998-1999: WHO/BTS summary report. Blood Transfusion Service, World Health Organization, Geneva, Switzerland.
Magda Anastassova Dineva*
Jean-Pierre Allain
Helen Lee
Department of Haematology
University of Cambridge/NBS
F40 EABC Site, Long Road
Cambridge CB2 2PT
United Kingdom

* Phone: 966502910024, Fax: 96635752255, E-mail: drpanhotra{at}gmail.com

Journal of Clinical Microbiology, December 2005, p. 6218-6219, Vol. 43, No. 12
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.12.6218-6219.2005





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