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 and studying virus replication. We developed positive- and negative-strand qRT-PCR assays to detect nsP3 of chikungunya virus (CHIKV), a positive-strand RNA alphavirus that causes epidemic fever, rash, and arthritis. The positive- and negative-strand qRT-PCR assays had limits of quantification of 1 and 3 log10 RNA copies/reaction, respectively. Compared to a published E1 diagnostic assay using 30 laboratory-confirmed clinical samples, the positive-strand nsP3 qRT-PCR assay had higher R(2) and efficiency and detected more positive samples. Peak viral load of 12.9 log(10) RNA copies/mL was reached on day 2 of illness, and RNA was detectable up to day 9, even in the presence of anti-CHIKV IgM. There was no correlation between viral load and persistent arthralgia. The positive-strand nsP3 assay is suitable for diagnosis, while the negative-strand 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
Descritores: Zika virus - Real Time PCR
Data de publicação: 2013
