Comparison of Polymerase Chain Reaction and Dot Hybridization with Enzyme‐Linked Immunoassay, Virological Examination and Polyacrylamide Gel Electrophoresis for the Detection of Porcine Rotavirus in Faecal Specimens

Winiarczyk, S.; Grądzki, Z.

doi:10.1046/j.1439-0450.1999.00288.x

Cited by 9 publications

(10 citation statements)

References 19 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This is in conformity with another study where Winiarczyk and Gradzki (1999) reported respective sensitivities and specificities of 100% and 94.7% for ELISA and 91.3% and 100% for PAGE for detection of porcine rotavirus in faecal samples.…”

Section: Resultssupporting

confidence: 92%

Comparative efficacy of immunological, molecular and culture assays for detection of group A rotavirus from faecal samples of buffalo (Bubalus bubalis) calves

Manuja

Prasad

Gulati

et al. 2010

Trop Anim Health Prod

View full text Add to dashboard Cite

Group A rotaviruses play an important role in causing gastroenteritis and mortality in buffalo (Bubalus bubalis) calves. A number of assays like RNA-polyacrylamide gel electrophoresis (RNA-PAGE), enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) and virus isolation have been employed for rotavirus diagnosis. We evaluated the comparative efficacy of different assays for detection of group A rotavirus in buffalo calves. A total of 455 faecal samples collected from five organized farms in northern India were screened by monoclonal antibody based ELISA, 33 (7.25%) samples were positive for group A rotavirus. The percent positivity ranged from 3.22% to 28% in different organized farms. The same samples were also tested by RNA-PAGE, which revealed classical 11 segments with 4:2:3:2 migration patterns in 14 faecal samples showing 3.08% positivity. Virus isolation was successfully done from 21 (4.61%) samples. However, only 15 (3.3%) samples yielded a specific product of 864 and 1,011 bp for VP4 and VP7 genes, respectively, by RT-PCR. The sensitivity and specificity of ELISA, RNA-PAGE and RT-PCR was 100%, 66.67% and 71.43% and 97%, 100% and 100%, respectively, considering virus isolation as standard test. ELISA being simple, fast and sensitive assay can be used as routine laboratory test for the diagnosis of group A rotavirus and field epidemiological studies.

show abstract

Section: Resultssupporting

confidence: 92%

Comparative efficacy of immunological, molecular and culture assays for detection of group A rotavirus from faecal samples of buffalo (Bubalus bubalis) calves

Manuja

Prasad

Gulati

et al. 2010

Trop Anim Health Prod

View full text Add to dashboard Cite

show abstract

“…Consequently, the sensitivity was proved to be slightly higher with the nested PCR than with the RLB assay in the analysis of clinical specimens. Those results are concordant with the finding of some researchers on the lesser sensitivity of the non-radioactive hybridization method in comparison with PCR amplification [Hopert et al, 1993;Winiarczyk and Gradzki, 1999;Myint, 2002]. However, the RLB has the advantage of the absence of possible false positives due to cross-over contamination.…”

Section: Discussionsupporting

confidence: 91%

Oligonucleotide array for simultaneous detection of respiratory viruses using a reverse‐line blot hybridization assay

Coiras

López-Huertas

López-Campos

et al. 2005

Journal of Medical Virology

View full text Add to dashboard Cite

The interest in developing new diagnostic methods based on arrays of multiple probes to detect and type simultaneously a wide range of different infectious agents is increasing. This becomes a necessity in the case of infectious agents such as respiratory viruses that cause diseases with very similar signs and symptoms. Such tools will permit rapid and accurate diagnosis of different agents causing respiratory infection leading to the most adequate prevention and/or treatment measures. In this article a reverse-line blot hybridization (RLB) assay for the detection of a wide range of respiratory viruses is presented and evaluated for its usefulness in routine diagnosis. This assay employs an array of 18 oligonucleotide probes immobilized on a nylon membrane. Biotin-labeled PCR products obtained with two multiplex reverse transcription (RT)-polymerase chain reaction (PCR) assays described previously, which allow for the detection of fourteen different groups of respiratory viruses, were hybridized to the oligonucleotide array. Detection was performed using a chemiluminescent method. The standardization of the method showed that the RLB assay could be an alternative to the nested PCR assay for enhancing the sensitivity in the detection of the amplified products, avoiding the problem of cross-over contamination, increasing the specificity, and therefore simplifying the method. This is of main interest in laboratories with few facilities. The feasibility and accuracy of the RT-PCR-RLB assay for detecting respiratory viruses proves that such approach could be a first stage to develop a microarray assay for routine diagnosis of infectious diseases.

show abstract

“…Other possible causative factors are the long-term storage of samples, which may decrease the amount of target viral genomes (Rådström et al 2002, Shay Fout et al 2003; the virus in question ma y belong to a different genotype from the ones for whi ch the primers used in the reactions were intended (Winiarczyk;Gradzki, 2002); or even the presence of non-group A rotaviruses circulating in the regions under study.…”

Section: Resultsmentioning

confidence: 99%

Molecular characterization of group A bovine rotavirus in southeastern and central-western Brazil, 2009-2010

et al. 2012

View full text Add to dashboard Cite

Rotavirus is an important cause of neonatal diarrhea in humans and several animal species, including calves. A study was conducted to examine 792 fecal samples collected from calves among 65 dairy and beef herds distributed in two of Brazil's major livestock producing regions, aiming to detect the occurrence of rotavirus and perform a molecular characterization of the rotavirus according to G and P genotypes in these regions. A total of 40 (5.05%) samples tested positive for rotavirus by the polyacrylamide gel electrophoresis (PAGE) technique. The molecular characterization was performed by multiplex semi-nested RT-PCR reactions, which indicated that the associations of genotypes circulating in herds in Brazil's southeastern region were G6P[11], G10P[11], G[-]P[5] + [11], G[-]P[6] in the state of São Paulo and G6P[11], G8P[5], G11P[11], G10P[11] in the state of Minas Gerais. In the central-western region, the genotypes G6P[5] + [11], G6P[5], G8P[-], G6P[11], G [-] P[1], G[-] P[11], and G[-] P[5] were detected in the state of Goiás, while the genotypes G6P[5], G8[P11], G6[P11], G8[P1], G8[P5], G6[P1] were circulating in herds in the state of Mato Grosso do Sul. The genotypic diversity of bovine rotavirus found in each region under study underlines the importance of characterizing the circulating samples in order to devise the most effective prophylactic measures.

show abstract

Comparison of Polymerase Chain Reaction and Dot Hybridization with Enzyme‐Linked Immunoassay, Virological Examination and Polyacrylamide Gel Electrophoresis for the Detection of Porcine Rotavirus in Faecal Specimens

Cited by 9 publications

References 19 publications

Comparative efficacy of immunological, molecular and culture assays for detection of group A rotavirus from faecal samples of buffalo (Bubalus bubalis) calves

Comparative efficacy of immunological, molecular and culture assays for detection of group A rotavirus from faecal samples of buffalo (Bubalus bubalis) calves

Oligonucleotide array for simultaneous detection of respiratory viruses using a reverse‐line blot hybridization assay

Molecular characterization of group A bovine rotavirus in southeastern and central-western Brazil, 2009-2010

Contact Info

Product

Resources

About