Real-time polymerase chain reaction for diagnosis and quantitation of negative strand of Chikungunya virus
Autor(es)Chiam Chun Wei, Chan Yoke Fun, Loong Shih Keng, Yong Sara Su Jin, Hooi Poh Sim, Sam I-Ching
ResumoQuantitative real-time polymerase chain reaction (qRT-PCR) is useful for diagnosis - studying virus replication. We developed positive- - negative-str- qRT-PCR assays to detect nsP3 of chikungunya virus (CHIKV), a positive-str- RNA alphavirus that causes epidemic fever, rash, - arthritis. The positive- - negative-str- qRT-PCR assays had limits of quantification of 1 - 3 log10 RNA copies/reaction, respectively. Compared to a published E1 diagnostic assay using 30 laboratory-confirmed clinical samples, the positive-str- nsP3 qRT-PCR assay had higher R(2) - efficiency - detected more positive samples. Peak viral load of 12.9 log(10) RNA copies/mL was reached on day 2 of illness, - RNA was detectable up to day 9, even in the presence of anti-CHIKV IgM. There was no correlation between viral load - persistent arthralgia. The positive-str- nsP3 assay is suitable for diagnosis, while the negative-str- nsP3 assay, which uses tagged primers to increase specificity, is useful for study of active viral replication kinetics.
Palavras-ChaveArthralgia; Chikungunya virus; Molecular diagnosis; Real-time PCR; Viral load
ImprentaDiagnostic Microbiology and Infectious Disease, v. 77, n. 2, p. 133-137, 2013
Identificador do Objeto Digital10.1016/j.diagmicrobio.2013.06.018
DescritoresChikungunya virus - Cell ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - Real Time PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Epidemic ; Chikungunya virus - Public health
Data de Publicação:2013