Strand-Specific Quantitative Reverse Transcription-Polymerase Chain Reaction Assay for Measurement of Arenavirus Genomic and Antigenomic RNAs.
Strand-Specific Quantitative Reverse Transcription-Polymerase Chain Reaction Assay for Measurement of Arenavirus Genomic and Antigenomic RNAs.
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DOI：
10.1371/journal.pone.0120043
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Botten J
中科院分区：
文献类型：
--
作者：
Haist K;Ziegler C;Botten J
Arenaviruses are bi-segmented, single-stranded RNA viruses that cause significant human disease. The manner in which they regulate the replication of their genome is not well-understood. This is partly due to the absence of a highly sensitive assay to measure individual species of arenavirus replicative RNAs. To overcome this obstacle, we designed a quantitative reverse transcription (RT)-PCR assay for selective quantitation of each of the lymphocytic choriomeningitis virus (LCMV) genomic or antigenomic RNAs. During the course of assay design, we identified a nonspecific priming phenomenon whereby, in the absence of an RT primer, cDNAs complementary to each of the LCMV replicative RNA species are generated during RT. We successfully circumvented this nonspecific priming event through the use of biotinylated primers in the RT reaction, which permitted affinity purification of primer-specific cDNAs using streptavidin-coated magnetic beads. As proof of principle, we used the assay to map the dynamics of LCMV replication at acute and persistent time points and to determine the quantities of genomic and antigenomic RNAs that are incorporated into LCMV particles. This assay can be adapted to measure total S or L segment-derived viral RNAs and therefore represents a highly sensitive diagnostic platform to screen for LCMV infection in rodent and human tissue samples and can also be used to quantify virus-cell attachment.
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影响因子：
5.4
作者：
EVANS, CF;BORROW, P;OLDSTONE, MBA
通讯作者：
OLDSTONE, MBA
DOI：
10.4269/ajtmh.1970.19.670
发表时间：
1970-01-01
影响因子：
3.3
作者：
FRAME, JD;BALDWIN, JM;TROUP, JM
通讯作者：
TROUP, JM
影响因子：
11.8
作者：
Emonet, Sebastien;Retornaz, Karine;Charrel, Remi N.
通讯作者：
Charrel, Remi N.
影响因子：
6.4
作者：
Kaiser, Philipp;Niederoest, Barbara;Fischer, Marek
通讯作者：
Fischer, Marek
影响因子：
3.1
作者：
Escaffre, O.;Queguiner, M.;Eterradossi, N.
通讯作者：
Eterradossi, N.