SARS-CoV infection was from at least two origins in the Taiwan area.
OBJECTIVE
Severe acute respiratory syndrome (SARS) is caused by a new coronavirus. Genomic sequence analysis will provide the molecular epidemiology and help to develop vaccines.


METHODS
We developed a rapid method to amplify and sequence the whole SARS-CoV genome from clinical specimens. The technique employed one-step multiplex RT-PCR to amplify the whole SARS-CoV genome, and then nested PCR was performed to amplify a 2-kb region separately. The PCR products were sequenced.


RESULTS
We sequenced the genomes of SARS-CoV from 3 clinical specimens obtained in Taiwan. The sequences were similar to those reported by other groups, except that 17 single nucleotide variations and two 2-nucleotide deletions, and a 1-nucleotide deletion were found. All the variations in the clinical specimens did not alter the amino acid sequence. Of these 17 sequenced variants, two loci (positions 26203 and 27812) were segregated together as a specific genotype - T:T or C:C. Phylogenetic analysis showed two major clusters of SARS patients in Taiwan.


CONCLUSIONS
We developed a very economical and rapid method to sequence the whole genome of SARS-CoV, which can avoid cultural influence. From our results, SARS patients in Taiwan may be infected from two different origins.
Shih
Mu-Chin
MC
Department of Molecular Medicine, China Medical University Hospital, Taichung, Taiwan.
Peck
Konan
K
Chan
Wen-Ling
WL
Chu
Yen-Ping
YP
Chen
Jui-Chang
JC
Tsai
Chang-Hai
CH
Chang
Jan-Gowth
JG
D000328:Adult; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D045169:Severe Acute Respiratory Syndrome / Q000453:epidemiology* / Q000821:virology*; D013624:Taiwan
D018076:DNA, Complementary; D012367:RNA, Viral