Molecular identification of mumps virus genotypes from clinical samples: standardized method of analysis.
A sensitive nested reverse transcription-PCR assay, targeting a short fragment of the gene encoding the small hydrophobic protein (SH gene), was developed to allow rapid characterization of mumps virus in clinical samples. The sensitivity and specificity of the assay were established using representative genotypes A, B, C, D, E, and F. Mumps virus RNA was characterized directly from cerebrospinal fluid (CSF) samples and in extracts of mumps virus isolates from patients with various clinical syndromes. Direct sequencing of products and subsequent phylogenetic analysis enabled genetic classification. A simple web-based system of sequence analysis was established. The study also allowed characterization of mumps virus strains from Argentina as part of a new subgenotype. This PCR assay for characterization of mumps infections coupled to a web-based analytical program provides a rapid method for identification of known and novel strains.
Palacios
G
G
Jerome L. and Dawn Greene Infectious Disease Laboratory, Mailman School of Public Health, Columbia University, 722 W 168th Street, Fl. 18, New York, NY 10032, USA. gp2050@columbia.edu
Jabado
O
O
Cisterna
D
D
de Ory
F
F
Renwick
N
N
Echevarria
J E
JE
Castellanos
A
A
Mosquera
M
M
Freire
M C
MC
Campos
R H
RH
Lipkin
W I
WI
D000595:Amino Acid Sequence; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D018792:Encephalitis, Viral / Q000821:virology; D005260:Female; D005838:Genotype; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008582:Meningitis, Aseptic / Q000821:virology; D008969:Molecular Sequence Data; D009107:Mumps / Q000821:virology; D009109:Mumps virus / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000235:genetics*
D012367:RNA, Viral; D014764:Viral Proteins; C067386:small hydrophobic protein, Mumps virus