scholarly journals Enzyme-Linked Immunosorbent Assay Based on Recombinant Human Group C Rotavirus Inner Capsid Protein (VP6) To Detect Human Group C Rotaviruses in Fecal Samples

1998 ◽  
Vol 36 (11) ◽  
pp. 3178-3181 ◽  
Author(s):  
V. L. A. James ◽  
P. R. Lambden ◽  
E. O. Caul ◽  
, and I. N. Clarke
Keyword(s):  
Immunosorbent Assay ◽  
Polyacrylamide Gel Electrophoresis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Group ◽  
Fecal Samples ◽  
Reverse Transcription Pcr ◽  
The United Kingdom ◽  
Group C Rotavirus ◽  
Group A ◽  
Rotavirus Infections

A recent study showed that 43% of a population in the United Kingdom were seropositive for group C rotavirus. The higher than expected incidence may be due to limited diagnosis of acute human group C rotavirus infections because no routine test is available. Human group C rotavirus infections are routinely diagnosed by electron microscopy (EM) and a negative group A rotavirus enzyme-linked immunosorbent assay (ELISA) result. An antigen-detection ELISA was developed with hyperimmune antibodies raised to human group C rotavirus recombinant VP6 (Bristol strain) expressed in insect cells. The assay was used to screen fecal samples to determine the prevalence of group C rotavirus infection. Samples positive by ELISA were confirmed by EM, polyacrylamide gel electrophoresis of double-stranded RNA, or detection of the VP6 gene by reverse transcription-PCR. Retrospective analysis indicated a 1 to 2% detection rate of positivity among samples from patients with acute diarrhea.

scholarly journals Seroepidemiology of Human Group C Rotavirus in Japan Based on a Blocking Enzyme-Linked Immunosorbent Assay

2001 ◽  
Vol 8 (1) ◽  
pp. 161-165 ◽  
Author(s):  
Mitsutaka Kuzuya ◽  
Ritsushi Fujii ◽  
Masako Hamano ◽  
Ritsuko Ohata ◽  
Hajime Ogura ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Age Groups ◽  
Neutralization Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Human Group ◽  
Serum Samples ◽  
Group C Rotavirus ◽  
Group A ◽  
Okayama Prefecture

ABSTRACT A novel blocking enzyme-linked immunosorbent assay (BL-ELISA) was developed for detection of antibodies to human group C rotavirus (CHRV). The specificity of the BL-ELISA was confirmed by using animal sera hyperimmunized to group A and group C rotaviruses and paired sera from five patients with acute CHRV gastroenteritis. Furthermore, there was concordance between the BL-ELISA and a neutralization assay for CHRV in 226 (95%) of 238 samples. By using the BL-ELISA, we determined the seroprevalence of CHRV in 704 serum samples obtained from nine different age groups of inhabitants of Okayama Prefecture, Japan, in 1992, 1994, and 1996. As a result, 211 sera (30%) were found to be positive for CHRV antibodies. The seroprevalence gradually increased with age and reached 52.7% in the oldest individuals. A further analysis of the youngest age group suggested that CHRVs predominantly prevail in persons older than 3 years of age in Japan. When comparing the three sampling years, a larger percentage of antibody-positive sera was detected in 1994 than in either 1992 or 1996 in individuals between 6 and 15 years of age, reflecting the occurrence of a CHRV outbreak among children during the winter of 1992 to 1993 that was previously documented. These results indicate that CHRV infections may occur more frequently in spite of the relatively low detection rate of the virus.

scholarly journals Seroepidemiology of Human Group C Rotavirus in South Africa

1999 ◽  
Vol 37 (12) ◽  
pp. 4142-4144 ◽  
Author(s):  
A. D. Steele ◽  
V. L. A. James
Keyword(s):  
South Africa ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Population ◽  
Human Group ◽  
South Africans ◽  
Group C Rotavirus ◽  
Percent Positivity ◽  
Rotavirus Infections ◽  
Rotavirus Vp6

Sera from three separate healthy population cohorts were used to determine the incidence of group C rotavirus infections in 1,356 South Africans. Using an enzyme-linked immunosorbent assay based on a recombinant group C rotavirus VP6 protein, the total percent positivity was found to be 34.4% (range, 33 to 38%), with almost half of the population infected after the age of 20 years.

scholarly journals Molecular Epidemiology of Human Group A Rotavirus Infections in the United Kingdom between 1995 and 1998

2000 ◽  
Vol 38 (12) ◽  
pp. 4394-4401 ◽  
Author(s):  
Miren Iturriza-Gómara ◽  
Jon Green ◽  
David W. G. Brown ◽  
Mary Ramsay ◽  
Ulrich Desselberger ◽  
...  
Keyword(s):  
United Kingdom ◽  
Molecular Epidemiology ◽  
Reverse Transcription ◽  
Animal Origin ◽  
Human Group ◽  
Group A Rotavirus ◽  
Reverse Transcription Pcr ◽  
The United Kingdom ◽  
Group A ◽  
Rotavirus Infections

The G and P types of 2,912 rotavirus-positive fecal specimens collected from eight geographical areas of the United Kingdom between 1995 and 1998 were determined by reverse transcription-PCR. Although 15 different G-P combinations were identified, G1P[8], G2P[4], G3P[8], and G4P[8] strains constituted 95% of all the rotaviruses typed. Other genotypes included G9P[6] and G9P[8], which were first identified in the United Kingdom in 1995, or other uncommon G and/or P types of strains that may have had an animal origin. Unusual combinations of G1 or G4 with P[4] and G2 with P[8] which may have arisen by reassortment between human strains were also identified. G1P[8] was the genotype most frequently found (57 to 87%) in each season, followed by G2P[4] in the 1995–1996 (18%) and 1997–1998 (16%) seasons, although the incidence of infection with this virus decreased significantly to 2% during the 1996–1997 season. Significant differences were seen in the distributions of G1P[8], G2P[4], and G9P[8] strains between children and adults, in the temporal distributions of G4P[8] and G9P[8] strains within a season, and in the geographical distributions of each of the four most common genotypes from one season to the next.

scholarly journals Characterization of Group C Rotaviruses Associated with Diarrhea Outbreaks in Feeder Pigs

1999 ◽  
Vol 37 (5) ◽  
pp. 1484-1488 ◽  
Author(s):  
Yunjeong Kim ◽  
Kyeong-Ok Chang ◽  
Barbara Straw ◽  
Linda J. Saif
Keyword(s):  
Amino Acid ◽  
Clinical Signs ◽  
Watery Diarrhea ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Group C Rotavirus ◽  
Group A Rotaviruses ◽  
Group A ◽  
Cowden Strain

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.

scholarly journals Dual Infection of Gnotobiotic Calves with Bovine Strains of Group A and Porcine-Like Group C Rotaviruses Influences Pathogenesis of the Group C Rotavirus

1999 ◽  
Vol 73 (11) ◽  
pp. 9284-9293 ◽  
Author(s):  
K. O. Chang ◽  
P. R. Nielsen ◽  
L. A. Ward ◽  
L. J. Saif
Keyword(s):  
Villous Atrophy ◽  
Polyacrylamide Gel Electrophoresis ◽  
Dual Infection ◽  
The United States ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Group C Rotavirus ◽  
Bovine Strain ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT There is serological evidence that bovine group C rotaviruses exist in the United States, but there are no reports of their isolation. Ninety fecal samples from calves with diarrhea, 81 samples from adult cows with diarrhea (winter dysentery), and 20 fecal samples from healthy adult cows were tested for group C rotaviruses by polyacrylamide gel electrophoresis, immune electron microscopy, and reverse transcription-PCR (RT-PCR). Three samples from adult cow diarrhea cases were positive only by RT-PCR, and a group C rotavirus was isolated from a positive sample in monkey kidney (MA104) cells (WD534tc/C). Genetically and serologically, the WD534tc/C strain was more closely related to the Cowden porcine group C strain than to the Shintoku bovine strain. Because the original cow feces also contained a group A rotavirus (detected after passage in cell culture), we hypothesized that such dual-rotavirus infections might play a role in the pathogenesis and host adaptation of rotaviruses. Thus, we examined the pathogenesis of WD534tc/C alone or combined with virulent (IND/A) or attenuated (NCDV/A) bovine group A rotaviruses in gnotobiotic calves. WD534tc/C alone induced diarrhea without (or with limited) virus shedding in inoculated calves (n = 3). In contrast, all calves coinfected with WD534tc/C and IND/A (n = 2) developed diarrhea and shed both viruses, whereas calves coinfected with WD534tc/C and NCDV/A (n= 3) developed diarrhea but did not shed either virus. Infection with WD534tc/C or NCDV/A alone caused only mild villous atrophy (jejunum and/or ileum), whereas dual infection with both viruses induced lesions throughout the small intestine. Although IND/A alone caused villous atrophy, more-widespread small intestinal lesions occurred in calves coinfected with WD534tc/C and IND/A. In conclusion, coinfection of calves with group A rotaviruses enhanced fecal shedding of a bovine group C rotavirus and the extent of histopathological lesions in the small intestines. Thus, our findings suggest a potential novel hypothesis involving dual infections for the adaptation of heterologous rotaviruses to new host species.

scholarly journals Fecal Cortisol and Progesterone Concentrations in Post Partus of Etawah Crossbreed Goat

2020 ◽  
Vol 151 ◽  
pp. 01031
Author(s):  
Claude M. Airin ◽  
Amelia Hana ◽  
Sarmin Sarmin ◽  
Pudji Astuti
Keyword(s):  
Immunosorbent Assay ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Stress Hormone ◽  
Fecal Samples ◽  
Fecal Cortisol ◽  
Fecal Extract ◽  
Last Stage ◽  
Preliminary Study ◽  
Freeze Dryer

Progesterone (P4) is a dominant hormone during pregnancy. In the later stage of pregnancy, the stress hormone particularly cortisol (C) may increase for initiating the parturition process as a consequence of fetal stress. This study was a preliminary study to compare the concentration of P4 and C in feces of Etawah Crossbreed Goat during their last stage of pregnancy and post partus. This study used 5 pregnant Etawah Crossbreed Goats (t 20th weeks) of pregnancy. Fecal samples were collected in the 20th week of pregnancy to 2 weeks of postpartum. All fecal samples were then dried using a freeze dryer (Labfreez FD10-MR) for 7 days at -80°C. Afterward, dried feces were pulverized and extracted by using 3ml of methanol 80%. The fecal extract was then analyzed the P4 and C concentrations using the enzyme-linked immunosorbent assay method. Concentrations of P4 and C metabolites in the last stage of pregnancy were 5,506.18 3,396.72 ng/g dry feces and 136,625.83 42,479.22 ng/g feces, respectively. Concentrations of P4 and C metabolites in the 2 weeks postpartum decreased at 669.38 P 643.9 ng/g feces and 110,295 / 14,378, 8 ng/g feces, respectively. It canbe concluded that there was a difference in the fecal progesterone and cortisol concentrations between the last phase of pregnancy and the postpartum phase.

scholarly journals Concomitant rotavirus serotypes 1 and 4 infections in a diarrhoeic child from Belém, Brazil

1996 ◽  
Vol 38 (4) ◽  
pp. 249-252 ◽  
Author(s):  
J.D.P. Mascarenhas ◽  
R.H.P. Gusmão ◽  
Y.B. Gabbay ◽  
T.A.F. Monteiro ◽  
J.B. Gomes ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Female Child ◽  
Polyacrylamide Gel ◽  
Current Concept ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rotavirus Vaccine ◽  
Rotavirus Infection ◽  
Diarrhoeic Child ◽  
Rotavirus Infections

Concomitant serotypes 1 and 4 infections were detected in a 15-month old female child with community-acquired diarrhoea which lasted 7 days and coursed with moderate dehydration. The evidence for dual rotavirus infection was offered by the following findings: a) enzyme-linked immunosorbent assay (ELISA) positive reactions to both 1 and 4 serotypes; and b) extra-migrating bands at electro-phoresis of RNA in polyacrylamide gel (PAGE). These results suggest that children living under poor sanitation conditions are heavily exposed to rotavirus infections; in addition, the co-circulation of different serotypes in the same setting sustains the current concept that a rotavirus vaccine should be rnultivalent, in order to protect children against the four epidemiologically important rotavirus G serotypes.

scholarly journals Assignment of Additional Anticapsular Antibody Concentrations to the Neisseria meningitidis Group A, C, Y, and W-135 Meningococcal Standard Reference Serum CDC1992

2002 ◽  
Vol 9 (3) ◽  
pp. 725-726 ◽  
Author(s):  
Cheryl M. Elie ◽  
Patricia K. Holder ◽  
Sandra Romero-Steiner ◽  
George M. Carlone
Keyword(s):  
Quality Control ◽  
Disease Control ◽  
Neisseria Meningitidis ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control And Prevention ◽  
Group A ◽  
Reference Serum ◽  
Centers For Disease Control

ABSTRACT We assigned additional enzyme-linked immunosorbent assay antibody concentrations (immunoglobulin G [IgG], IgM, and IgA, and total) to the Neisseria meningitidis standard reference serum CDC1992 for groups Y and W-135 to 12 Centers for Disease Control and Prevention quality control sera. These assignments will supplement previous assignments and will aid in the evaluation of present and developing vaccines.

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