scholarly journals A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA

1994 ◽  
Vol 6 (2) ◽  
pp. 175-181 ◽  
Author(s):  
A. Lucchelli ◽  
S. Y. Kang ◽  
M. K. Jayasekera ◽  
A. V. Parwani ◽  
D. H. Zeman ◽  
...  

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.

1993 ◽  
Vol 111 (2) ◽  
pp. 407-412 ◽  
Author(s):  
Y. Pongsuwanna ◽  
K. Taniguchi ◽  
F. Wakasugi ◽  
Y. Sutivijit ◽  
M. Chiwakul ◽  
...  

SummaryA total of 241 group A rotavirus-positive stool samples collected from diarrhoeic patients in Thailand between July 1988 and June 1991 were characterized for their serotypes by enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies and by a polymerase chain reaction (PCR). In July 1988–June 1989, serotype 1 was the most prevalent (63·4%), followed by serotype 4 (11·0%) and serotype 2 (8·5%). In July 1989–June 1990, 59·8% were serotype 1, 24·3% were serotype 2, and 6·1 % were serotype 3. In contrast, in July 1990–June 1991, serotype 3 was detected in the highest frequency (40·5%), 29·9% were serotype 1, and 27·3% were serotype 2. Thus, a distinct yearly change of serotype distribution of rotavirus in Thailand was observed in the three consecutive years. In particular, it was of note that the prevalence of serotype 3 greatly increased, in contrast to the previous studies in which almost no serotype 3 rotaviruses were detected in the years 1983–8 in Thailand.


Virology ◽  
1992 ◽  
Vol 191 (1) ◽  
pp. 272-281 ◽  
Author(s):  
Hiroshi Tsunemitsu ◽  
Clem K. Ojeh ◽  
Baoming Jiang ◽  
Ron A. Simkins ◽  
Peggy A. Weilnau ◽  
...  

1989 ◽  
Vol 102 (3) ◽  
pp. 523-530 ◽  
Author(s):  
S. K. Ghosh ◽  
T. N. Naik

SUMMARYMonoclonal antibodies specific for rotavirus subgroup 1 (SG1) and subgroup 2 (SG2) were used to analyse by enzyme immunoassay (EIA) the subgroups of human rotavirus isolates obtained from three different parts of India during the period September 1985 to July 1987. We identified one isolate which failed to react with either SG1 or SG2 specific monoclonal antibodies, although it reacted well with a monoclonal antibody specific for group A rotaviruses. This finding suggests that it belongs to a new rotavirus subgroup. Further, another isolate was found to belong to SG1 although it had a ‘long’ electropherotype.


2000 ◽  
Vol 7 (2) ◽  
pp. 288-292 ◽  
Author(s):  
Yousif Al-Yousif ◽  
Fahad Al-Majhdi ◽  
Cindy Chard-Bergstrom ◽  
Joe Anderson ◽  
Sanjay Kapil

ABSTRACT Hybridomas secreting monoclonal antibodies (MAbs) against the Nebraska calf diarrhea strain of bovine rotavirus (BRV) were characterized. Indirect fluorescent-antibody assay, immunodot assay, and immunoprecipitation were used to select hybridomas that produced anti-BRV MAbs. Seven of the MAbs were shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot assay to be reactive with the BRV outer capsid protein, VP7, which has a molecular mass of 37.5 kDa. None of the seven MAbs were reactive with canine rotavirus, bovine coronavirus, or uninfected Madin-Darby bovine kidney cells. Two clones, 8B4 (immunoglobulin G2a [IgG2a]) and 2B11 (IgG1), were found suitable for use in an antigen capture enzyme-linked immunosorbent assay for detecting BRV in bovine fecal samples. Both were subtype A specific (G6 subtype) but did not react with all isolates of BRV group A.


1999 ◽  
Vol 37 (5) ◽  
pp. 1484-1488 ◽  
Author(s):  
Yunjeong Kim ◽  
Kyeong-Ok Chang ◽  
Barbara Straw ◽  
Linda J. Saif

Feces and serum specimens were collected from three farms in Michigan on which ∼50-lb (8- to 9-week-old) pigs experienced diarrhea just after placement into all-in-all-out finishing barns. The clinical signs (profuse watery diarrhea lasting about 2 weeks and no vomiting) were similar on all farms, and the morbidity rate was high (ranging from 60 to 80%) but without mortality. Eleven diarrheic fecal samples from the farms were tested for group A and C rotaviruses by immune electron microscopy (IEM) and various assays. IEM indicated that the fecal samples reacted only with antiserum against group C rotaviruses, and polyacrylamide gel electrophoresis indicated that the samples had characteristic genomic electropherotypes for group C rotavirus. Group C rotavirus was detected by cell culture immunofluorescence (CCIF) tests in nine fecal samples, but no group A rotavirus was detected by enzyme-linked immunosorbent assay or CCIF. By reverse transcription (RT)-PCR, all 11 fecal samples were positive for group C rotaviruses, with only 2 samples positive for group A rotaviruses. However, a second amplification of RT-PCR products using nested primers detected group A rotaviruses in all samples. Analysis of nucleotide and deduced amino acid sequences of the RT-PCR product (partial-length VP7) of the group C rotavirus showed 87.2 to 91% nucleotide identity and 92.6 to 95.9% amino acid identity among two strong samples from the different farms and the Cowden strain of porcine group C rotavirus. All nine convalescent-phase serum samples tested had neutralizing antibodies to the Cowden strain, and the majority of them had neutralizing antibody against group A rotaviruses (OSU or/and Gottfried strains) by fluorescent focus neutralization tests. Although group C rotaviruses have been reported as a cause of sporadic diarrhea in suckling or weanling pigs, to our knowledge, this is the first report of epidemic diarrhea outbreaks associated with group C rotavirus in older pigs.


Author(s):  
M. De Beer ◽  
I. Peenze ◽  
V.M. Da Costa Mendes ◽  
A.D. Steele

The performance characteristics of 2 enzyme immunoassays (ELISAs) and 4 latex agglutination assays (LXs) were evaluated for the detection of bovine rotavirus in faecal specimens of young calves with diarrhoea. A total of 26 specimens from calves less than 5 months of age were examined with different commercial assays and compared with electron microscopy (EM) as the gold standard and with polyacrylamide gel electrophoresis (PAGE) for the detection of atypical, non-group A rotaviruses. In the 2nd study, EIA (Dako) and LX (Murex), the assays of choice, were used to analyse 97 further faecal specimens from calves with diarrhoea. The ELISAs proved to be the most sensitive compared with the other tests used. The EM and PAGE are 100 % specific although slightly less sensitive than the commercial assays. The results show that all the commercial assays can accurately detect rotavirus in the stools of calves with gastroenteritis, although the suitability and choice of assay will depend upon the requirements of individual laboratories.


2019 ◽  
Vol 31 (1) ◽  
pp. 87-92

Rotaviruses are regarded as the most common cause of viral gastroenteritis and are responsible for considerable morbidity and mortality among children especially under five years of age worldwide. In developing countries like Myanmar, where diarrhoea is in the priority childhood disease, rotavirus surveillance and detection of rotavirus genotypes are utmost important. A hospital-based, cross-sectional descriptive study was conducted at Yangon Children‟s Hospital among under five children admitted for acute diarrhoea from January to October 2016. This study includes detection of Group A rotavirus antigen by commercial enzyme-linked immunosorbent assay (ELISA) and genotyping by multiplex RT-PCR. From a total of 488 collected samples, rotavirus antigen was detected in 219 samples (45%). Rotavirus diarrhoea was most common among the age of 6-11 months (38.8%) followed by 12-23 months (37.9%). The results showed that boys were more commonly affected than girls. Detection of rotavirus positivity was peak in February (57.6 %). Out of 219 stool samples with positive ELISA result, 40 stool samples with high optical density value were proceeded for further determination of G and P genotypes. Regarding distribution of G genotypes, the most common G genotype was G9 which comprised 45%, and that of P genotype was P[8] which comprised 92.5%. Regarding combination of G and P genotypes, the most frequent combination is G9P[8], and it constituted 42.5%. Untypable genotypes were seen in 30% of G and 2.5% of P typing. As rotavirus infection can be prevented by vaccine, WHO recommended that rotavirus vaccination should be included in national immunization program especially in countries where prevalence of rotavirus is high. The distribution of G and P genotypes is important in consideration of appropriate vaccine in pre-vaccination and evaluation of effectiveness of vaccine in post-vaccination period. Therefore, the information on currently circulating genotypes of rotavirus in this study will serve as valuable data for vaccination programme.


Pathogens ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 959
Author(s):  
Celeste M. Donato ◽  
Julie E. Bines

Group A rotaviruses belong to the Reoviridae virus family and are classified into G and P genotypes based on the outer capsid proteins VP7 and VP4, respectively [...]


Sign in / Sign up

Export Citation Format

Share Document