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. 1999 Aug;37(8):2706-8.
doi: 10.1128/JCM.37.8.2706-2708.1999.

Identification of human rotavirus strains with the P[14] genotype by PCR

Affiliations

Identification of human rotavirus strains with the P[14] genotype by PCR

S Arista et al. J Clin Microbiol. 1999 Aug.

Abstract

A seminested PCR typing assay has been extended to identify rotavirus strains with the P[14] genotype. The specificity of the method was confirmed by Southern hybridization and by restriction analysis with the enzyme AluI. One out of four human rotavirus (HRV) strains with unusual subgroup-electropherotype linkage but none out of 50 HRV strains with usual linkage was typed as P[14].

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Figures

FIG. 1
FIG. 1
Electrophoretic analysis of PCR products of HRV reference strains. Lane 1, DNA molecular weight marker VIII (a mixture of pUCBM21 DNA HpaII and DraI-plus-HindIII digest; Boehringer Mannheim); lane 2, strain Wa (G1, P[8]); lane 3, strain DS1 (G2, P[4]); lane 4, strain ST3 (G4, P[6]); lane 5, strain AU-1 (G3, P[9]); lane 6, strain PA169 (G6, P[14]). (A) Ethidium bromide-stained gel showing RT-PCR products after second amplification, visualized under UV light. (B) Results of Southern hybridization with biotinylated HRV P[14] genotype-specific PR-1 probe.
FIG. 2
FIG. 2
Electrophoresis profiles of PCR-amplified gene 4 cDNA of strain PA169 (lanes 2 and 3) and of fecal isolate PA-5/89 (lanes 4 and 5), before and after, respectively, digestion with AluI, visualized by ethidium bromide staining of the gel. Lane 1, DNA molecular weight marker VIII (Boehringer Mannheim).

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