Abstract
The presence of hepatitis B virus DNA in the sera of individuals is the most definitive marker of an active viral infection. We have used polymerase chain reaction detection of hepatitis B virus DNA directly on whole blood dried as a spot on filter paper. The method is rapid, specific, and sensitive and has the ability to detect as little as 10 virus particles by ethidium bromide staining of the polymerase chain reaction-amplified products. The method is cost-effective, and the stability of the spots makes the collection and transportation of potentially infectious blood safe.
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- Beebe J. L., Briggs L. C. Evaluation of enzyme-linked immunoassay systems for detection of human immunodeficiency virus type 1 antibody from filter paper disks impregnated with whole blood. J Clin Microbiol. 1990 Apr;28(4):808–810. doi: 10.1128/jcm.28.4.808-810.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brechot C., Hadchouel M., Scotto J., Degos F., Charnay P., Trepo C., Tiollais P. Detection of hepatitis B virus DNA in liver and serum: a direct appraisal of the chronic carrier state. Lancet. 1981 Oct 10;2(8250):765–768. doi: 10.1016/s0140-6736(81)90182-3. [DOI] [PubMed] [Google Scholar]
- Cassol S., Salas T., Arella M., Neumann P., Schechter M. T., O'Shaughnessy M. Use of dried blood spot specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction. J Clin Microbiol. 1991 Apr;29(4):667–671. doi: 10.1128/jcm.29.4.667-671.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheyrou A., Guyomarc'h C., Jasserand P., Blouin P. Improved detection of HBV DNA by PCR after microwave treatment of serum. Nucleic Acids Res. 1991 Jul 25;19(14):4006–4006. doi: 10.1093/nar/19.14.4006. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ganem D., Varmus H. E. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. doi: 10.1146/annurev.bi.56.070187.003251. [DOI] [PubMed] [Google Scholar]
- Garrick M. D., Dembure P., Guthrie R. Sickle-cell anemia and other hemoglobinopathies. Procedures and strategy for screening employing spots of blood on filter paper as specimens. N Engl J Med. 1973 Jun 14;288(24):1265–1268. doi: 10.1056/NEJM197306142882403. [DOI] [PubMed] [Google Scholar]
- Hoofnagle J. H., Schafer D. F. Serologic markers of hepatitis B virus infection. Semin Liver Dis. 1986 Feb;6(1):1–10. doi: 10.1055/s-2008-1040787. [DOI] [PubMed] [Google Scholar]
- Kain K. C., Lanar D. E. Determination of genetic variation within Plasmodium falciparum by using enzymatically amplified DNA from filter paper disks impregnated with whole blood. J Clin Microbiol. 1991 Jun;29(6):1171–1174. doi: 10.1128/jcm.29.6.1171-1174.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaneko S., Miller R. H., Feinstone S. M., Unoura M., Kobayashi K., Hattori N., Purcell R. H. Detection of serum hepatitis B virus DNA in patients with chronic hepatitis using the polymerase chain reaction assay. Proc Natl Acad Sci U S A. 1989 Jan;86(1):312–316. doi: 10.1073/pnas.86.1.312. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
- Landers T. A., Greenberg H. B., Robinson W. S. Structure of hepatitis B Dane particle DNA and nature of the endogenous DNA polymerase reaction. J Virol. 1977 Aug;23(2):368–376. doi: 10.1128/jvi.23.2.368-376.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Larzul D., Chevrier D., Thiers V., Guesdon J. L. An automatic modified polymerase chain reaction procedure for hepatitis B virus DNA detection. J Virol Methods. 1990 Jan;27(1):49–60. doi: 10.1016/0166-0934(90)90145-6. [DOI] [PubMed] [Google Scholar]
- McCabe E. R., McCabe L. Screening for PKU in sick or premature neonates. J Pediatr. 1983 Sep;103(3):502–503. doi: 10.1016/s0022-3476(83)80451-x. [DOI] [PubMed] [Google Scholar]
- Walter E., Blum H. E., Offensperger W. B., Zeschnigk C., Offensperger S., Gerok W. Spot-blot hybridization assay for the detection of hepatitis B virus DNA in serum: factors determining its sensitivity and specificity. Hepatology. 1987 May-Jun;7(3):557–562. doi: 10.1002/hep.1840070323. [DOI] [PubMed] [Google Scholar]
- Yokosuka O., Omata M., Hosoda K., Tada M., Ehata T., Ohto M. Detection and direct sequencing of hepatitis B virus genome by DNA amplification method. Gastroenterology. 1991 Jan;100(1):175–181. doi: 10.1016/0016-5085(91)90598-f. [DOI] [PubMed] [Google Scholar]