Cloning of Human Picobirnavirus Genomic Segments and Development of an RT-PCR Detection Assay

Abstract

Nearly full-length genomic segments 2 and a partial-length genomic segment 1 of human picobirnavirus were cloned and sequenced. The clones were derived from viruses obtained from human immunodeficiency virus (HIV)-infected patients in Atlanta, Georgia (strains 3-GA-91 and 4-GA-91) and a nonHIV-infected person from China (strain 1-CHN-97). The picobirnavirus genomic segments lacked sequence similarities with other viral sequences in GenBank and EMBL. Comparison of genomic segment 1 from a human and a rabbit picobirnavirus identified a region of 127 nucleotides with 54.7% identity. The genomic segments 2 of the 4-GA-91 and 1-CHN-97 strains had 41.4% nucleic acid identity and 30.0% amino acid similarity and contained amino acid motifs typical of RNA-dependent RNA polymerase genes. Reverse transcription-PCR detection assays were developed with primers targeted to the genomic segments 2 of strains 4-GA-91 or 1-CHN-97. Picobirnaviruses related to the China strain were the predominant viruses detected in stool samples from people in four countries on three continents. Picobirnaviruses were detected in samples from two outbreaks of gastroenteritis in long-term elder care facilities but were not determined to be the primary pathogen. Our findings support the view that picobirnaviruses constitute a distinct family of viruses.

Detection and Molecular Characterization of Porcine Picobirnavirus in Feces of Domestic Pigs from Kolkata, India

“…In this study, we carried out the detection and molecular characterization assays for porcine PBV in feces of asymptomatic domestic pigs in Kolkata, India. Out of 11 samples, two showed very faint PBV positive (large genome profile) by PAGE and both the samples showed positive for genogroup I PBV by RT-PCR using genogroup specific primer pair developed by Rosen et al [35]. Further characterization by sequencing and phylogenetic analyses revealed that the genogroup I porcine PBV strain detected during this study clustered with hitherto reported porcine, murine and human PBV strains reported from various countries.…”

“…PBVs were detected from immunocompromised patients [24] in which implied to be ''opportunistic pathogens'' and have also been detected from asymptomatic hosts [11]. The question whether PBV is an 'innocuous agent' of the intestine remains to be investigated [2,7,19,31,35].…”

“…The reverse transcription-polymerase chain reaction (RT-PCR) protocol developed by Rosen et al [35] enabled the detection and molecular characterization of PBV studies, worldwide. With the two sets of primer pairs that specifically amplify small fragments within the RdRp gene of segment 2, PBV strains have been classified into two genogroups (genogroup I with 201 bp amplicon and genogroup II with 369 bp amplicon) represented by prototype strains 1-CHN-97 and 4-GA-91 isolated in China and USA, respectively.…”

“…The primer pairs described by Rosen et al [35] were used for molecular characterization experiments. RT-PCR was carried out following the protocol of Bhattacharya et al [3].…”

Genogroup I picobirnaviruses in pigs: evidence for genetic diversity and relatedness to human strains

“…PBVs have also been detected in patients with gastroenteritis. PBVs have been frequently detected as coinfections together with rotaviruses, caliciviruses and astroviruses (Bányai et al, 2003;Bhattacharya et al, 2006aBhattacharya et al, , b, 2007Rosen et al, 2000). In addition, the higher detection rates of PBVs in immuncompromised patients without the detection of conventional enteric pathogens (Giordano et al, 1998(Giordano et al, , 1999Gonzalez et al, 1998;Grohmann et al, 1993;Martinez et al, 2003) suggest that PBV might be an opportunistic pathogen.…”