scholarly journals Generation of Infectious Recombinant Human Rotaviruses from Just 11 Cloned cDNAs Encoding the Rotavirus Genome

2019 ◽  
Vol 93 (8) ◽  
Author(s):  
Satoshi Komoto ◽  
Saori Fukuda ◽  
Masanori Kugita ◽  
Riona Hatazawa ◽  
Chitose Koyama ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Human Group ◽  
Severe Diarrhea ◽  
Generation Vaccine ◽  
Single Animal ◽  
Growth Properties ◽  
Group A Rotaviruses ◽  
Group A ◽  
Foreign Genes ◽  
Reassortant Viruses

ABSTRACTThe generation of recombinant group A rotaviruses (RVAs) entirely from cloned cDNAs has been described only for a single animal RVA strain, simian SA11-L2. We recently developed an optimized RVA reverse genetics system based on only RVA cDNAs (11-plasmid system), in which the concentration of cDNA plasmids containing the NSP2 and NSP5 genes is 3- or 5-fold increased in relation to that of the other plasmids. Based on this approach, we generated a recombinant human RVA (HuRVA)-based monoreassortant virus containing the VP4 gene of the simian SA11-L2 virus using the 11-plasmid system. In addition to this monoreassortant virus, authentic HuRVA (strain KU) was also generated with the 11-plasmid system with some modifications. Our results demonstrate that the 11-plasmid system involving just RVA cDNAs can be used for the generation of recombinant HuRVA and recombinant HuRVA-based reassortant viruses.IMPORTANCEHuman group A rotavirus (HuRVA) is a leading pathogen causing severe diarrhea in young children worldwide. In this paper, we describe the generation of recombinant HuRVA (strain KU) from only 11 cloned cDNAs encoding the HuRVA genome by reverse genetics. The growth properties of the recombinant HuRVA were similar to those of the parental RVA, providing a powerful tool for better understanding of HuRVA replication and pathogenesis. Furthermore, the ability to manipulate the genome of HuRVAs “to order” will be useful for next-generation vaccine production for this medically important virus and for the engineering of clinical vectors expressing any foreign genes.

scholarly journals Survey of Human Group C Rotaviruses in Japan during the Winter of 1992 to 1993

1998 ◽  
Vol 36 (1) ◽  
pp. 6-10 ◽  
Author(s):  
Mitsutaka Kuzuya ◽  
Ritsushi Fujii ◽  
Masako Hamano ◽  
Masao Yamada ◽  
Kuniko Shinozaki ◽  
...  
Keyword(s):  
Large Scale ◽  
Blot Hybridization ◽  
Human Group ◽  
Dot Blot ◽  
Passive Hemagglutination ◽  
Group A Rotaviruses ◽  
Group A ◽  
Outer Capsid ◽  
Very High ◽  
Vp7 Gene

Fecal specimens from patients with acute diarrhea were collected from 10 prefectures in Japan over a 6-month period (November 1992 to April 1993), and the specimens that were negative for human group A rotaviruses were screened for the presence of human group C rotaviruses (CHRVs) by the reverse passive hemagglutination test. Of 784 specimens examined, 53 samples (6.8%) that were collected in 7 of 10 prefectures were positive for CHRV, indicating that CHRVs are widely distributed across Japan. Most of the CHRV isolates were detected in March and April, and CHRVs mainly prevailed in children ages 3 to 8 years. The genome electropherotypes of eight strains isolated in five individual prefectures were surprisingly similar to each other and were different from those of CHRV strains isolated to date. The outer capsid glycoprotein (VP7) gene homologies of the isolates retrieved in 1993 were subsequently analyzed by the dot blot hybridization method. As a result, the VP7 genes of the isolates revealed very high levels of homology not only with each other but also with the VP7 gene of the OK118 strain isolated in 1988. These results suggest that a large-scale outbreak of CHRV occurred during the winter of 1992 and 1993 in Japan.

scholarly journals Genotypic determination of human group A rotaviruses from Goa and Meghalaya states, India

Heliyon ◽  
2020 ◽  
Vol 6 (8) ◽  
pp. e04521
Author(s):  
Abhay Raorane ◽  
Zunjar Dubal ◽  
Sandeep Ghatak ◽  
Michael Mawlong ◽  
B. Susngi ◽  
...  
Keyword(s):  
Human Group ◽  
Group A Rotaviruses ◽  
Group A

scholarly journals Detection and Genotyping of Human Group A Rotaviruses by Oligonucleotide Microarray Hybridization

2002 ◽  
Vol 40 (7) ◽  
pp. 2398-2407 ◽  
Author(s):  
V. Chizhikov ◽  
M. Wagner ◽  
A. Ivshina ◽  
Y. Hoshino ◽  
A. Z. Kapikian ◽  
...  
Keyword(s):  
Oligonucleotide Microarray ◽  
Microarray Hybridization ◽  
Human Group ◽  
Group A Rotaviruses ◽  
Group A

scholarly journals Group B rotaviruses similar to strain CAL-1, have been circulating in Western India since 1993

2004 ◽  
Vol 132 (4) ◽  
pp. 745-749 ◽  
Author(s):  
S. D. KELKAR ◽  
J. K. ZADE
Keyword(s):  
Severe Disease ◽  
Polyacrylamide Gel Electrophoresis ◽  
Epidemiological Studies ◽  
Western India ◽  
Human Group ◽  
Older Children ◽  
Pcr Products ◽  
Group A Rotaviruses ◽  
Group A ◽  
Group B

Generally, group A rotaviruses are the most common cause of paediatric diarrhoea. However, group B rotavirus, adult diarrhoea rotavirus (ADRV), was found to be involved in epidemics of severe gastroenteritis in several areas of China during 1982–1983 and had resulted in more than one million cases among adults as well as older children. Human group B rotavirus has been rarely reported outside China, but has been detected first from five adults with diarrhoea in Kolkata, India during 1997–1998 (strain CAL-1). During epidemiological studies at the National Institute of Virology (NIV) on hospitalized diarrhoea patients at Pune, India, faecal specimens from patients of >5 years age, which were negative for group A rotavirus by ELISA were tested by polyacrylamide gel electrophoresis (PAGE). We detected rotavirus RNA migration patterns similar to that of group B rotavirus in three faecal specimens from adults, two from the specimens collected in 1993 and one in 1998 from sporadic diarrhoea cases. RT–PCR was carried out using primers derived from gene 8 which codes for the NS2 protein, followed by nested PCR, which confirmed the presence of group B rotavirus in all three specimens. The sequences of the PCR products of NIV specimens were compared with that of CAL-1, ADRV and IDIR (infectious diarrhoea of infant rat) belonging to group B rotaviruses. The sequence analysis of the PCR products showed the highest identity with CAL-1, which was reported from Kolkata, India during 1997–1998. The finding suggests that human group B rotaviruses have been circulating in Pune, India, since 1993. This emerging virus may lead to more severe disease among adults in India. There is a need for surveillance of group B rotavirus infections, especially in adult diarrhoea cases and seroepidemiological studies on group B rotavirus are required among humans and animals of Western Maharashtra, India.

Changing pattern of human group A rotaviruses: Emergence of G12 as an important pathogen among children in eastern India

2006 ◽  
Vol 36 (3) ◽  
pp. 183-188 ◽  
Author(s):  
S. Samajdar ◽  
V. Varghese ◽  
P. Barman ◽  
S. Ghosh ◽  
U. Mitra ◽  
...  
Keyword(s):  
Eastern India ◽  
Human Group ◽  
Group A Rotaviruses ◽  
Group A

scholarly journals Transfection of exogenous rotavirus rearranged RNA segments in cells infected with a WT rotavirus results in subsequent gene rearrangements

2014 ◽  
Vol 95 (9) ◽  
pp. 2089-2098 ◽  
Author(s):  
Sarah Duponchel ◽  
Cécile Troupin ◽  
Lan Trang Vu ◽  
Aurélie Schnuriger ◽  
Germain Trugnan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Acute Gastroenteritis ◽  
Reverse Genetics ◽  
Gene Rearrangements ◽  
Recombinant Viruses ◽  
Bovine Strain ◽  
The Family ◽  
Viral Progeny ◽  
Group A Rotaviruses ◽  
Group A

Group A rotaviruses, members of the family Reoviridae, are a major cause of infantile acute gastroenteritis. The rotavirus genome consists of 11 dsRNA segments. In some cases, an RNA segment is replaced by a rearranged RNA segment, which is derived from its standard counterpart by partial sequence duplication. It has been shown that some rearranged segments are preferentially encapsidated into viral progenies after serial passages in cell culture. Based on this characteristic, a reverse genetics system was used previously to introduce exogenous segment 7 rearrangements into an infectious rotavirus. This study extends this reverse genetics system to RNA segments 5 and 11. Transfection of exogenous rotavirus rearranged RNA segment 5 or 11 into cells infected with a WT helper rotavirus (bovine strain RF) resulted in subsequent gene rearrangements in the viral progeny. Whilst recombinant viruses were rescued with an exogenous rearranged segment 11, the exogenous segment was modified by a secondary rearrangement. The occurrence of spontaneous rearrangements of WT or exogenous segments is a major hindrance to the use of this reverse genetics approach.

scholarly journals Clinical Symptoms of Human Rotavirus Infection Observed in Children in Sokoto, Nigeria

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
B. R. Alkali ◽  
A. I. Daneji ◽  
A. A. Magaji ◽  
L. S. Bilbis
Keyword(s):  
Developing Countries ◽  
Clinical Symptoms ◽  
Primary Data ◽  
Human Group ◽  
Infantile Diarrhoea ◽  
Elisa Kit ◽  
Urban Hospitals ◽  
Human Rotavirus ◽  
Group A Rotaviruses ◽  
Group A

Rotavirus has been identified among the most important causes of infantile diarrhoea, especially in developing countries. The present study was undertaken to determine the occurrence and clinical symptoms of human rotavirus disease among children presenting with varying degree of diarrhoea in selected urban hospitals in Sokoto metropolis, Nigeria. Diarrhoea samples were collected from 200 diarrheic children younger than 5 years of age and tested using a commercially available DAKO Rotavirus ELISA kit which detects the presence of human group A rotaviruses. A questionnaire, based on WHO generic protocol, was completed for each child to generate the primary data. Of the total number of samples collected, 51 were found to be positive for human group A rotavirus indicating 25.5% prevalence of the disease in Sokoto state. The symptoms associated with the disease were analyzed and discussed.

scholarly journals Rapid generation of rotavirus single-gene reassortants by means of eleven plasmid-only based reverse genetics

2020 ◽  
Vol 101 (8) ◽  
pp. 806-815 ◽  
Author(s):  
Saori Fukuda ◽  
Riona Hatazawa ◽  
Yoshiki Kawamura ◽  
Tetsushi Yoshikawa ◽  
Takayuki Murata ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Single Gene ◽  
Basic Research ◽  
Clinical Applications ◽  
Phenotypic Traits ◽  
Selection Processes ◽  
Group A Rotaviruses ◽  
Group A ◽  
Specific Restriction ◽  
Rapid Generation

Reassortment is an important mechanism in the evolution of group A rotaviruses (RVAs), yielding viruses with novel genetic and phenotypic traits. The classical methods for generating RVA reassortants with the desired genetic combinations are laborious and time-consuming because of the screening and selection processes required to isolate a desired reassortant. Taking advantage of a recently developed RVA reverse genetics system based on just 11 cloned cDNAs encoding the RVA genome (11 plasmid-only system), we prepared a panel of simian SA11-L2 virus-based single-gene reassortants, each containing 1 segment derived from human KU virus of the G1P[8] genotype. It was shown that there was no gene-specific restriction of the reassortment potential. In addition to these 11 single-gene reassortants, a triple-gene reassortant with KU-derived core-encoding VP1–3 gene segments with the SA11-L2 genetic background, which make up a virion composed of the KU-based core, and SA11-L2-based intermediate and outer layers, could also be prepared with the 11 plasmid-only system. Finally, for possible clinical application of this system, we generated a series of VP7 reassortants representing all the major human RVA G genotypes (G1–4, G9 and G12) efficiently. The preparation of each of these single-gene reassortants was achieved within just 2 weeks. Our results demonstrate that the 11 plasmid-only system allows the rapid and reliable generation of RVA single-gene reassortants, which will be useful for basic research and clinical applications.

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