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
Resumo: Quantitative 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-Chave: Arthralgia; Chikungunya virus; Molecular diagnosis; Real-time PCR; Viral load
Imprenta: Diagnostic Microbiology and Infectious Disease, v. 77, n. 2, p. 133-137, 2013
Identificador do objeto digital: 10.1016/j.diagmicrobio.2013.06.018
Descritores: Chikungunya 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
