Genotyping of recent measles virus strains from Russia and Vietnam by nucleotide-specific multiplex PCR.
The nucleoprotein genes of 49 measles virus (MV) strains circulating in Russia between 2000 and 2006 and in Vietnam in 2003 were analyzed by genotype-specific PCR and the results were compared with their sequences. The sequences revealed the presence of genotypes H1 and H2 in the center (Nha Trang) and the north (Hanoi) of Vietnam, respectively. The relative diversity of the H2 strains suggested an endemic circulation of these viruses in the capital. In contrast genotype H1 strains from Nha Trang were homogenous genetically, which may indicate a recent importation. The strains obtained from 12 different regions of the Russian Federation were assigned to the genotypes H1, D4, and D6. Most strains (81.4%) were correctly genotyped by a multiplex PCR method which was sensitive to genotype-specific mutations [Kremer et al. (2004): J Clin Microbiol 42: 3017-3022]. Ambiguous or negative results for some clade H and genotype D6 strains were due to point mutations in the type-specific primer binding sites. After exchanging a single nucleotide in both the clade H- and the genotype D6-specific primers, all strains were assigned correctly to their genotype. A simplified procedure for use in Vietnam was developed to distinguish directly between genotypes H1 and H2 and any non-H genotype. These results demonstrate that our multiplex PCR method can be adapted easily to new sequence variants or specific epidemiological situations, and thus be very useful for rapid genotyping of large number of samples even in laboratories which do not have sequencing facilities.
Kremer
Jacques R
JR
Institute of Immunology and WHO Collaborative Centre for Measles and WHO European Regional Reference Laboratory for Measles and Rubella, Laboratoire National de Santé, L-1011, Luxembourg.
Nguyen
Giang H
GH
Shulga
Sergey V
SV
Nguyen
Phuc H
PH
Nguyen
Ut T
UT
Tikhonova
Nina T
NT
Muller
Claude P
CP
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D005838:Genotype; D006801:Humans; D008457:Measles / Q000453:epidemiology / Q000821:virology*; D008459:Measles virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D017720:Molecular Epidemiology; D019590:Nucleocapsid Proteins / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012426:Russia / Q000453:epidemiology; D014744:Vietnam / Q000453:epidemiology
D017931:DNA Primers; D004279:DNA, Viral; D019590:Nucleocapsid Proteins