Genotyping of human bocavirus using a restriction length polymorphism.
Sequencing analysis of the isolates of a recently identified pathogen associated with respiratory infections, human bocavirus (HBoV), allowed for identification of two virus genotypes of the virus. In the current article a new method for a simple and fast differentiation of HBoV genotypes in clinical materials is described. The test includes an amplification of a 309 bp fragment of VP1/VP2 gene of HBoV from nasopharyngeal aspirates with a subsequent incubation of a PCR mix with the BstAPI endonuclease. Upon such a digestion, the DNA fragment derived from the genotype I HBoV isolates forms two fragments of 150 and 159 bp, while that obtained from genotype 2 isolates remains unrestricted. The developed technique may be used in epidemiological studies of HBoV infection and analysis of the potential differences in biological characteristics of HboV genotypes.
Ditt
Vanessa
V
Institute for Virology, University of Bonn, Sigmund-Freud-Strasse 25, 53105 Bonn, Germany.
Viazov
Sergei
S
Tillmann
Ramona
R
Schildgen
Verena
V
Schildgen
Oliver
O
D001483:Base Sequence; D052660:Bocavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment
D004279:DNA, Viral