Abstract:
:A simple, sensitive, and specific polymerase chain reaction (PCR) protocol for the detection of human immunodeficiency virus type 1 (HIV-1) is described. We have improved all three PCR steps: sample preparation, DNA amplification, and detection of the amplified product. Some of the improvements have been described previously, but they have never been combined into a complete PCR protocol. Peripheral blood mononuclear cells were lysed directly in a buffer containing sodium dodecyl sulfate, Triton X-100, and proteinase K. This crude cell lysate was amplified in a two-step PCR, first with outer primers and then with inner primers nested within the first primers. The PCR product was visualized by agarose gel electrophoresis and ethidium bromide staining. Thus, we avoided conventional DNA extraction as well as hybridization for the detection of the PCR product. The samples were analyzed with four sets of nested primers (JA4 through JA7, JA9 through JA12, JA13 through JA16, and JA17 through JA20) designed to amplify HIV-1 gag, env gp120, env gp41, and pol sequences, respectively. We were able to amplify HIV-1 sequences in all samples from 90 HIV-1-seropositive individuals with mostly mild symptoms. Of these individuals, 24 were negative in HIV-1 isolation and 9 were selected because they were infected by African and Haitian HIV-1 strains. Eighty-five (94%) individuals were positive with at least three of four primer sets. Samples from 26 healthy blood donors, as well as cells infected in vitro with human immunodeficiency virus type 2 and human T-cell leukemia virus type I, were negative in PCR, thus demonstrating the specificity of the amplification.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Albert J,Fenyö EMdoi
10.1128/JCM.28.7.1560-1564.1990subject
Has Abstractpub_date
1990-07-01 00:00:00pages
1560-4issue
7eissn
0095-1137issn
1098-660Xjournal_volume
28pub_type
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.06469-11
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01128-06
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.19.4.538-540.1984
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.5.1150-1153.1995
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.4.672-674.1986
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.7.1181-1185.1987
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.1.1.15-24.1975
更新日期:1975-01-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.32.9.2221-2224.1994
更新日期:1994-09-01 00:00:00
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.36.2.526-530.1998
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pub_type: 杂志文章
doi:10.1128/JCM.36.8.2346-2348.1998
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.10.2175-2179.1989
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.22.4.488-489.1985
更新日期:1985-10-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/JCM.38.7.2756-2759.2000
更新日期:2000-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2014-06-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2006-02-01 00:00:00
abstract::The DNA relatedness and restriction fragment length polymorphism patterns of whole-cell DNA and the general phenotypic properties of 14 isolates of the form species Candida parapsilosis representing three diverse genetic groups were compared. The data confirm the unrelatedness of the three groups at the species level....
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.1.216-218.1998
更新日期:1998-01-01 00:00:00