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Detection and quantitation of HTLV-1 and HTLV-2 mRNA species by real-time RT-PCR.

Abstract：

:HTLV-1 and HTLV-2 are highly related delta-retroviruses that infect and transform T-lymphocytes, but have distinct pathogenic properties. HTLV replication and survival requires the expression of multiple gene products from an unspliced and a series of highly related alternatively spliced mRNA species. To date, the comparative levels of all known HTLV-1 and HTLV-2 viral mRNAs in different transformed cell lines and at different stages of virus infection have not been assessed. In this study, we compiled a series of oligonucleotide primer pairs and probes to quantify both HTLV-1 and HTLV-2 mRNA species using real-time RT-PCR. The optimized reaction for detection of each mRNA had amplification efficiency greater than 90% with a linear range spanning 25-2.5 x 10(7) copies. The R(2)'s of all standard curves were greater than 0.97. Quantitation of HTLV mRNAs between different cell lines showed variability (gag/pol>or=tax/rex>env>or=accessory proteins), but the overall levels of each mRNA relative to each other within a cell line were similar. These results provide a method to quantify all specific mRNAs from both HTLV-1 and HTLV-2, which can be used to evaluate further viral gene expression and correlate transcript levels to key stages of the virus life cycle and ultimately, pathogenesis.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Li M,Green PL

doi

10.1016/j.jviromet.2007.01.023

subject

Has Abstract

pub_date

2007-06-01 00:00:00

pages

159-68

issue

1-2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(07)00049-3

journal_volume

142

pub_type

杂志文章

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