Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

Gehad Elkady; Jie Zhu; Qingjie Peng; Ming Chen; Xin Liu; Yingyu Chen; Changmin Hu; Huanchun Chen; Aizhen Guo

doi:10.1016/j.meegid.2021.104715

Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

Gehad Elkady, Jie Zhu, Qingjie Peng, Ming Chen, Xin Liu, Yingyu Chen, Changmin Hu, Huanchun Chen, Aizhen Guo

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Abstract

Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.

Original language	English (US)
Article number	104715
Journal	Infection, Genetics and Evolution
Volume	89
DOIs	https://doi.org/10.1016/j.meegid.2021.104715
State	Published - Apr 2021
Externally published	Yes

Bibliographical note

Funding Information:
This study was supported by The National Key Research and Development Program of China (# 2018YFD0501802 ), the Special Fund for Chinese Agricultural Research System (Beef/yaks) ( #CARS-37 ), and the Special Fund for National Distinguished Scholars in Agricultural Research and Technical Innovative Team.

Publisher Copyright:
© 2021 The Author(s)

Keywords

Bovine rotavirus
Calf diarrhea
ESI-LC-MS/MS genome constellation
Isolation
RVA
RVB

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.meegid.2021.104715

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@article{ddc2737b546441d5b3e713e097623654,

title = "Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves",

abstract = "Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.",

keywords = "Bovine rotavirus, Calf diarrhea, ESI-LC-MS/MS genome constellation, Isolation, RVA, RVB",

author = "Gehad Elkady and Jie Zhu and Qingjie Peng and Ming Chen and Xin Liu and Yingyu Chen and Changmin Hu and Huanchun Chen and Aizhen Guo",

note = "Funding Information: This study was supported by The National Key Research and Development Program of China (# 2018YFD0501802 ), the Special Fund for Chinese Agricultural Research System (Beef/yaks) ( #CARS-37 ), and the Special Fund for National Distinguished Scholars in Agricultural Research and Technical Innovative Team. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

month = apr,

doi = "10.1016/j.meegid.2021.104715",

language = "English (US)",

volume = "89",

journal = "Infection, Genetics and Evolution",

issn = "1567-1348",

publisher = "Elsevier",

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TY - JOUR

T1 - Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

AU - Elkady, Gehad

AU - Zhu, Jie

AU - Peng, Qingjie

AU - Chen, Ming

AU - Liu, Xin

AU - Chen, Yingyu

AU - Hu, Changmin

AU - Chen, Huanchun

AU - Guo, Aizhen

N1 - Funding Information: This study was supported by The National Key Research and Development Program of China (# 2018YFD0501802 ), the Special Fund for Chinese Agricultural Research System (Beef/yaks) ( #CARS-37 ), and the Special Fund for National Distinguished Scholars in Agricultural Research and Technical Innovative Team. Publisher Copyright: © 2021 The Author(s)

PY - 2021/4

Y1 - 2021/4

N2 - Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.

AB - Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.

KW - Bovine rotavirus

KW - Calf diarrhea

KW - ESI-LC-MS/MS genome constellation

KW - Isolation

KW - RVA

KW - RVB

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DO - 10.1016/j.meegid.2021.104715

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AN - SCOPUS:85099374520

SN - 1567-1348

VL - 89

JO - Infection, Genetics and Evolution

JF - Infection, Genetics and Evolution

M1 - 104715

ER -

Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves

Abstract

Bibliographical note

Keywords

UN SDGs

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