title: SARS-CoV infection was from at least two origins in the Taiwan area.
abstract: 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.
authors.affiliation: Department of Molecular Medicine, China Medical University Hospital, Taichung, Taiwan.
authors.forename: Mu-Chin
authors.initials: MC
authors.lastname: Shih
authors.forename: Konan
authors.initials: K
authors.lastname: Peck
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Chan
authors.forename: Yen-Ping
authors.initials: YP
authors.lastname: Chu
authors.forename: Jui-Chang
authors.initials: JC
authors.lastname: Chen
authors.forename: Chang-Hai
authors.initials: CH
authors.lastname: Tsai
authors.forename: Jan-Gowth
authors.initials: JG
authors.lastname: Chang
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral