Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction.
A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein E1 (E1) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354- and 294-base pair (bp) cDNA fragments were amplified effectively from the nsP1 and E1 genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial E1 gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype.
Department of Virology, Institute of Tropical Medicine, Nagasaki University, Sakamoto, Nagasaki, Japan. rainbow@net.nagasaki-u.ac.jp
F
F
Hasebe
M C
MC
Parquet
B D
BD
Pandey
E G M
EG
Mathenge
K
K
Morita
V
V
Balasubramaniam
Z
Z
Saat
A
A
Yusop
M
M
Sinniah
S
S
Natkunam
A
A
Igarashi
D000330:Aedes; D018354:Alphavirus Infections / Q000175:diagnosis* / Q000453:epidemiology* / Q000821:virology; D000818:Animals; D002478:Cells, Cultured; D002646:Chikungunya virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004196:Disease Outbreaks; D005838:Genotype; D006801:Humans; D008296:Malaysia / Q000453:epidemiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins