Abstract:
:The aim of this study was the development of a real-time PCR for HIV DNA quantification in whole blood leucocytes providing an alternative assay to those already described, almost based on the gag gene detection. The technique (pbs-rtPCR assay) is more rapid (the whole assay required less than 5h), easy to perform, omitting both PBMC purification step and data normalization to a housekeeping gene, when compared to previously published assays. Our method is able to detect all group M HIV-1 subtypes in the highly conserved primer-binding site (PBS) region and to avoid the interference by retroviral endogenous sequences in HIV DNA quantification. The sensitivity was 100% for 2 copies even in the presence of high amounts of genomic DNA (1 microg). To monitor the HIV DNA level in all possible clinical conditions, the assay has been validated and compared with a previously developed gag-PCR assay on 73 HIV-1 HAART-treated patients with a plasma HIV-1 RNA range from <50 to >500,000 copies/ml. The 50% of the samples with a viremia below the limit of detection (50 copies/ml) was found to contain HIV DNA between 2 and 10 copies/microg DNA. The pbs-rtPCR offers a significant improvement in the percentage of quantitatively detectable sample (99%) compared with the gag-PCR (42%) suggesting caution in the choice of HIV DNA assay.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Casabianca A,Gori C,Orlandi C,Forbici F,Federico Perno C,Magnani Mdoi
10.1016/j.mcp.2007.05.005subject
Has Abstractpub_date
2007-10-01 00:00:00pages
368-78issue
5-6eissn
0890-8508issn
1096-1194pii
S0890-8508(07)00040-0journal_volume
21pub_type
杂志文章abstract::Research has been focused on the detection of Mycobacterium avium subspecies paratuberculosis (MAP) in pasteurized milk; however, pasteurized milk is a key ingredient in a variety of food products. Therefore, MAP contamination in milk-derived products must be investigated. We undertook a six-month study to investigate...
journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
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