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Int J Mol Epidemiol Genet 2010;2(1):61-72.
Original Article
Detection and molecular characterization of multiple strains of Picobirnavirus
causing mixed infection in a diarrhoeic child: emergence of prototype
genogroup II-like strain in Kolkata, India
Balasubramanian Ganesh, Shigeo Nagashima, Souvik Ghosh, Seegekote M. Nataraju, Krishnan Rajendran, Byomkesh
Manna, Thandavarayan Ramamurthy, Swapan K. Niyogi, Suman Kanungo, Dipika Sur, Nobumichi Kobayashi, Triveni Krishnan
Division of Virology, Division of Data Management, Division of Microbiology, Division of Epidemiology, National
Institute of Cholera and Enteric Diseases (NICED), P-33, C.I.T. Road, Scheme-XM, Beliaghata, Kolkata 700 010, West
Bengal, India. Department of Hygiene, Sapporo Medical University School of Medicine, S-1 W-17, Chuo-Ku, Sapporo
060-8556, Japan.
Received September 27, 2010; accepted December 20, 2010; Epub December 27, 2010; published January 1, 2011
Abstract: Background: Picobirnaviruses (PBVs) associated with viral gastroenteritis were reported from humans and
several animal species to date. PBVs belonging to family Picobirnaviridae under proposed order Diplornavirales are
small, non-enveloped, with bisegmented dsRNA genome. Methods: PBV was detected by polyacrylamide gel electrophoresis
(PAGE) and silver staining. Confirmatory RT-PCR using primer pair PicoB25 (+) and PicoB43 (-) for genogroup
I PBV and PicoB23(+) and PicoB24(-) for genogroup II PBV, resulted in amplicons of 201bp and 369bp respectively.
The amplicons of genogroup I PBV were cloned and sequenced; amplicon of genogroup II PBV was directly sequenced.
Further, the phylogenetic relationship and genetic diversity of strains from Kolkata was compared with hitherto
reported PBV strains. Results: In PAGE, a faecal specimen showed three sets of PBV with large profile bisegmented
genomic RNA with slight variation in migration pattern. Molecular cloning experiments confirmed that PBV/
Human/INDIA/GPBV6/2007 had mixed infection comprising four different strains of PBV genogroup I [GPBV6C1PGPBV6C4P]
and one PBV genogroup II strain [GPBV6G2P]. Conclusion: Sequence comparison and phylogenetic
analysis of gene segment 2 of GPBV6 clones (C1, C2, C3 and C4) revealed low nucleotide identities (59-63%) and
distant genetic relatedness to other human and porcine genogroup I picobirnaviruses. The strain GPBV6G2P represents
another PBV genogroup II strain after prototype strain 4-GA-91/USA as genogroup II PBVs have seldom been
reported to date, except from Kolkata, India and Netherlands. We are reporting the first incidence of detection of
multiple strain (mixed) infection of picobirnavirus [genogroups I and II] from a diarrhoeic child in a slum community of
Kolkata, India. (IJMEG1009004).
Keywords: Genogroup I and II Picobirnavirus, bisegmented dsRNA virus, viral diarrhea, mixed infection
Full Text PDF
Address all correspondence to:
Dr. Triveni Krishnan,
Division of Virology, National Institute of Cholera and Enteric Diseases (NICED)
P-33, C.I.T. Road, Scheme-XM, Beliaghata, Kolkata 700 010
West Bengal, India.
Tel 91-33-23633852; Fax 91-33-23632398
E-mail: drtriveni.krishnan@gmail.com
Original Article
Detection and molecular characterization of multiple strains of Picobirnavirus
causing mixed infection in a diarrhoeic child: emergence of prototype
genogroup II-like strain in Kolkata, India
Balasubramanian Ganesh, Shigeo Nagashima, Souvik Ghosh, Seegekote M. Nataraju, Krishnan Rajendran, Byomkesh
Manna, Thandavarayan Ramamurthy, Swapan K. Niyogi, Suman Kanungo, Dipika Sur, Nobumichi Kobayashi, Triveni Krishnan
Division of Virology, Division of Data Management, Division of Microbiology, Division of Epidemiology, National
Institute of Cholera and Enteric Diseases (NICED), P-33, C.I.T. Road, Scheme-XM, Beliaghata, Kolkata 700 010, West
Bengal, India. Department of Hygiene, Sapporo Medical University School of Medicine, S-1 W-17, Chuo-Ku, Sapporo
060-8556, Japan.
Received September 27, 2010; accepted December 20, 2010; Epub December 27, 2010; published January 1, 2011
Abstract: Background: Picobirnaviruses (PBVs) associated with viral gastroenteritis were reported from humans and
several animal species to date. PBVs belonging to family Picobirnaviridae under proposed order Diplornavirales are
small, non-enveloped, with bisegmented dsRNA genome. Methods: PBV was detected by polyacrylamide gel electrophoresis
(PAGE) and silver staining. Confirmatory RT-PCR using primer pair PicoB25 (+) and PicoB43 (-) for genogroup
I PBV and PicoB23(+) and PicoB24(-) for genogroup II PBV, resulted in amplicons of 201bp and 369bp respectively.
The amplicons of genogroup I PBV were cloned and sequenced; amplicon of genogroup II PBV was directly sequenced.
Further, the phylogenetic relationship and genetic diversity of strains from Kolkata was compared with hitherto
reported PBV strains. Results: In PAGE, a faecal specimen showed three sets of PBV with large profile bisegmented
genomic RNA with slight variation in migration pattern. Molecular cloning experiments confirmed that PBV/
Human/INDIA/GPBV6/2007 had mixed infection comprising four different strains of PBV genogroup I [GPBV6C1PGPBV6C4P]
and one PBV genogroup II strain [GPBV6G2P]. Conclusion: Sequence comparison and phylogenetic
analysis of gene segment 2 of GPBV6 clones (C1, C2, C3 and C4) revealed low nucleotide identities (59-63%) and
distant genetic relatedness to other human and porcine genogroup I picobirnaviruses. The strain GPBV6G2P represents
another PBV genogroup II strain after prototype strain 4-GA-91/USA as genogroup II PBVs have seldom been
reported to date, except from Kolkata, India and Netherlands. We are reporting the first incidence of detection of
multiple strain (mixed) infection of picobirnavirus [genogroups I and II] from a diarrhoeic child in a slum community of
Kolkata, India. (IJMEG1009004).
Keywords: Genogroup I and II Picobirnavirus, bisegmented dsRNA virus, viral diarrhea, mixed infection
Full Text PDF
Address all correspondence to:
Dr. Triveni Krishnan,
Division of Virology, National Institute of Cholera and Enteric Diseases (NICED)
P-33, C.I.T. Road, Scheme-XM, Beliaghata, Kolkata 700 010
West Bengal, India.
Tel 91-33-23633852; Fax 91-33-23632398
E-mail: drtriveni.krishnan@gmail.com
