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. 1997 Jan;35(1):278–280. doi: 10.1128/jcm.35.1.278-280.1997

Comparison of in-house and commercial sample preparation and PCR amplification systems for detection of human immunodeficiency virus type 1 DNA in blood samples from Tanzanian adults.

E Lyamuya 1, U Bredberg-Rådén 1, J Albert 1, O Grankvist 1, V Msangi 1, C Kagoma 1, F Mhalu 1, G Biberfeld 1
PMCID: PMC229556  PMID: 8968925

Abstract

This study compared the performance of several in-house nested PCR systems and the Amplicor human immunodeficiency virus type 1 (HIV-1) PCR kit in the detection of HIV-1 DNA in Tanzanian samples prepared by two different methods. All six of the in-house primer sets evaluated had a higher sensitivity for HIV DNA detection in samples prepared by the Amplicor PCR sample preparation method than in those prepared by the Ficoll-Isopaque (FIP) density gradient centrifugation method. A sensitivity of 100% was achieved by combining two in-house primer sets. The sensitivity of the standard Amplicor HIV-1 PCR kit was only 59%, whereas a modified Amplicor HIV-1 PCR test had a sensitivity of 98%. Our data show that Tanzanian samples prepared by the Amplicor preparation method are more suitable for HIV-1 PCR testing than samples prepared by the FIP method. The modified, but not the standard, Amplicor HIV-1 PCR kit provides an alternative to the nested in-house PCR technique for the diagnosis of HIV infection.

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Selected References

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  1. Albert J., Fenyö E. M. Simple, sensitive, and specific detection of human immunodeficiency virus type 1 in clinical specimens by polymerase chain reaction with nested primers. J Clin Microbiol. 1990 Jul;28(7):1560–1564. doi: 10.1128/jcm.28.7.1560-1564.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borkowsky W., Krasinski K., Pollack H., Hoover W., Kaul A., Ilmet-Moore T. Early diagnosis of human immunodeficiency virus infection in children less than 6 months of age: comparison of polymerase chain reaction, culture, and plasma antigen capture techniques. J Infect Dis. 1992 Sep;166(3):616–619. doi: 10.1093/infdis/166.3.616. [DOI] [PubMed] [Google Scholar]
  3. Bredberg-Rådén U., Urassa E., Grankvist O., Massawe A., Lyamuya E., Kawo G., Msemo G., Kazimoto T., Mgone J., Mbena E. Early diagnosis of HIV-1 infection in infants in Dar es Salaam, Tanzania. Clin Diagn Virol. 1995 Aug;4(2):163–173. doi: 10.1016/0928-0197(95)00010-6. [DOI] [PubMed] [Google Scholar]
  4. Brinchmann J. E., Albert J., Vartdal F. Few infected CD4+ T cells but a high proportion of replication-competent provirus copies in asymptomatic human immunodeficiency virus type 1 infection. J Virol. 1991 Apr;65(4):2019–2023. doi: 10.1128/jvi.65.4.2019-2023.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clewley J. P. The polymerase chain reaction, a review of the practical limitations for human immunodeficiency virus diagnosis. J Virol Methods. 1989 Aug;25(2):179–187. doi: 10.1016/0166-0934(89)90031-1. [DOI] [PubMed] [Google Scholar]
  6. Eisenstein B. I. The polymerase chain reaction. A new method of using molecular genetics for medical diagnosis. N Engl J Med. 1990 Jan 18;322(3):178–183. doi: 10.1056/NEJM199001183220307. [DOI] [PubMed] [Google Scholar]
  7. Fransen K., van Kerckhoven I., Piot P., van der Groen G. Evaluation and comparison of the Amplicor HIV-1 PCR kit with an 'in-house' nested PCR. Clin Diagn Virol. 1995 Dec;4(4):311–319. doi: 10.1016/0928-0197(95)00016-x. [DOI] [PubMed] [Google Scholar]
  8. Grankvist O., Gustafsson A., Bredberg-Rådén U., Pallangyo K., Mhalu F., Biberfeld P., Biberfeld G., Wadell G. Selection of primers of optimal sensitivity for the detection of HIV-1 from Africa and Europe by polymerase chain reaction. AIDS. 1991 May;5(5):575–578. [PubMed] [Google Scholar]
  9. Jackson J. B., Drew J., Lin H. J., Otto P., Bremer J. W., Hollinger F. B., Wolinsky S. M. Establishment of a quality assurance program for human immunodeficiency virus type 1 DNA polymerase chain reaction assays by the AIDS Clinical Trials Group. ACTG PCR Working Group, and the ACTG PCR Virology Laboratories. J Clin Microbiol. 1993 Dec;31(12):3123–3128. doi: 10.1128/jcm.31.12.3123-3128.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Khadir A., Coutlée F., Saint-Antoine P., Olivier C., Voyer H., Kessous-Elbaz A. Clinical evaluation of Amplicor HIV-1 test for detection of human immunodeficiency virus type 1 proviral DNA in peripheral blood mononuclear cells. J Acquir Immune Defic Syndr Hum Retrovirol. 1995 Jul 1;9(3):257–263. [PubMed] [Google Scholar]
  11. 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]
  12. Loche M., Mach B. Identification of HIV-infected seronegative individuals by a direct diagnostic test based on hybridisation to amplified viral DNA. Lancet. 1988 Aug 20;2(8608):418–421. doi: 10.1016/s0140-6736(88)90412-6. [DOI] [PubMed] [Google Scholar]
  13. Louwagie J., Janssens W., Mascola J., Heyndrickx L., Hegerich P., van der Groen G., McCutchan F. E., Burke D. S. Genetic diversity of the envelope glycoprotein from human immunodeficiency virus type 1 isolates of African origin. J Virol. 1995 Jan;69(1):263–271. doi: 10.1128/jvi.69.1.263-271.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Louwagie J., McCutchan F. E., Peeters M., Brennan T. P., Sanders-Buell E., Eddy G. A., van der Groen G., Fransen K., Gershy-Damet G. M., Deleys R. Phylogenetic analysis of gag genes from 70 international HIV-1 isolates provides evidence for multiple genotypes. AIDS. 1993 Jun;7(6):769–780. doi: 10.1097/00002030-199306000-00003. [DOI] [PubMed] [Google Scholar]
  15. Lynch C. E., Madej R., Louie P. H., Rodgers G. Detection of HIV-1 DNA by PCR: evaluation of primer pair concordance and sensitivity of a single primer pair. J Acquir Immune Defic Syndr. 1992;5(5):433–440. [PubMed] [Google Scholar]
  16. Ou C. Y., Kwok S., Mitchell S. W., Mack D. H., Sninsky J. J., Krebs J. W., Feorino P., Warfield D., Schochetman G. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science. 1988 Jan 15;239(4837):295–297. doi: 10.1126/science.3336784. [DOI] [PubMed] [Google Scholar]
  17. Saag M. S., Hahn B. H., Gibbons J., Li Y., Parks E. S., Parks W. P., Shaw G. M. Extensive variation of human immunodeficiency virus type-1 in vivo. Nature. 1988 Aug 4;334(6181):440–444. doi: 10.1038/334440a0. [DOI] [PubMed] [Google Scholar]
  18. Scarlatti G., Lombardi V., Plebani A., Principi N., Vegni C., Ferraris G., Bucceri A., Fenyö E. M., Wigzell H., Rossi P. Polymerase chain reaction, virus isolation and antigen assay in HIV-1-antibody-positive mothers and their children. AIDS. 1991 Oct;5(10):1173–1178. doi: 10.1097/00002030-199110000-00003. [DOI] [PubMed] [Google Scholar]
  19. Sison A. V., Campos J. M. Laboratory methods for early detection of human immunodeficiency virus type 1 in newborns and infants. Clin Microbiol Rev. 1992 Jul;5(3):238–247. doi: 10.1128/cmr.5.3.238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Siwka W., Schwinn A., Baczko K., Pardowitz I., Mhalu F., Shao J., Rethwilm A., ter Meulen V. vpu and env sequence variability of HIV-1 isolates from Tanzania. AIDS Res Hum Retroviruses. 1994 Dec;10(12):1753–1754. doi: 10.1089/aid.1994.10.1753. [DOI] [PubMed] [Google Scholar]
  21. Vandamme A. M., Fransen K., Debaisieux L., Marissens D., Sprecher S., Vaira D., Vandenbroucke A. T., Verhofstede C. Standardisation of primers and an algorithm for HIV-1 diagnostic PCR evaluated in patients harbouring strains of diverse geographical origin. The Belgian AIDS Reference Laboratories. J Virol Methods. 1995 Feb;51(2-3):305–316. doi: 10.1016/0166-0934(94)00126-2. [DOI] [PubMed] [Google Scholar]
  22. Whetsell A. J., Drew J. B., Milman G., Hoff R., Dragon E. A., Adler K., Hui J., Otto P., Gupta P., Farzadegan H. Comparison of three nonradioisotopic polymerase chain reaction-based methods for detection of human immunodeficiency virus type 1. J Clin Microbiol. 1992 Apr;30(4):845–853. doi: 10.1128/jcm.30.4.845-853.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zazzi M., Romano L., Brasini A., Valensin P. E. Low human immunodeficiency virus titer and polymerase chain reaction false-negatives. J Infect Dis. 1992 Apr;165(4):779–780. doi: 10.1093/infdis/165.4.779. [DOI] [PubMed] [Google Scholar]

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