Comparison of agarose gel electrophoresis and RNA-PAGE for rapid detection of rotavirus from faecal samples

Dubal, Z. B.; Mawlong, Michael; Susngi, B.; Sanjukta, Rajkumari; Puro, K.; Ghatak, Sandeep; Sen, Alaattin; Shakuntala, I.; Barbuddhe, Sukhadeo B.; Ahuja, Anuj; Bhattacharjee, Uttaran

doi:10.1080/09712119.2014.896262

Cited by 4 publications

(3 citation statements)

References 19 publications

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“…The mixed infections found in this study population were P [8]P [10]. The P [6] and P [9] genotypes were not found in circulation during this study period.…”

Section: Rotavirus Infection and Socio-demographic Factorsmentioning

confidence: 57%

“…Once solidified, the agarose gel was placed into the gel box (electrophoresis unit) and fill the gel box with 1xTBE until the gel is covered, about 3 µl of gel loading dye was added to each amplicon and gently mix, ladder was carefully loaded into the centre lane of the gel and samples into the additional wells of the gel; the gel was run at 80-150 V for 30 min; The power was turned off, the electrodes disconnected from the power source, and the gel carefully removed from the gel box. UVP Photodoc-It Imaging System (Fisher Scientific) was used to visualize the DNA fragments referred as 'bands' due to their appearance on the gel alongside 100 base pair ladders (Dubai et al 2015).…”

Section: Loading Of Samples and Running Of Agarose Gelmentioning

confidence: 99%

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Molecular Characterization and Risk Factors Associated with Rotavirus among Children with diarrhoea.

Demekong,

Onu,

Okorie

et al. 2024

Preprint

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Introduction: Rotavirus-induced acute diarrhoea continues to pose a significant health threat to children under 5 years old worldwide, despite the availability of effective vaccines. This study aimed to assess the prevalence, associated risk factors, and circulating genotypes of rotavirus among children under 5 admitted to Ikom Medical Centre due to diarrhea. Methodology: Stool samples were collected from 294 children under 5 with diarrhoea at Ikom Medical Centre. Rotavirus antigen detection was performed using rapid chromatographic immunoassay. Molecular techniques such as polyacrylamide gel electrophoresis (PAGE), RT-PCR, and semi-nested multiplex RT-PCR were used to analyze rotavirus strains. Data were obtained through questionnaires, and statistical significance was assessed using the chi-square test (P < 0.05). Results: Among the 294 samples, 20.4% (60/294) tested positive for rotavirus. Significant risk factors influencing rotavirus prevalence included feeding status, toilet system, and duration of diarrhoea. The most prevalent genotype for VP7 was G4 (26.7%), and for VP4, it was genotype P[4] (23.3%). Predominant genotype combinations were G4P [4] (11.6%) and G12P[4] (6.7%), while non-typeable P[NT] was observed in 41.7% of cases. Conclusion: This study highlights the circulation of rotavirus in Ikom, Cross River State, emphasizing its neglected status despite its significant global impact on child mortality. Urgent attention is needed to address rotavirus as a serious public health concern, particularly in low-resource settings.

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“…The mixed infections found in this study population were P [8]P [10]. The P [6] and P [9] genotypes were not found in circulation during this study period.…”

Section: Rotavirus Infection and Socio-demographic Factorsmentioning

confidence: 57%

Section: Loading Of Samples and Running Of Agarose Gelmentioning

confidence: 99%

Molecular Characterization and Risk Factors Associated with Rotavirus among Children with diarrhoea.

Demekong,

Onu,

Okorie

et al. 2024

Preprint

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“…Many sensitive and fast molecular techniques have been used for detection of viral agents causing gastroenteritis and screening of stool samples to give an indication of the true occurrence of the causative agent of acute gastroenteritis in the pediatric patients [10]. Virus genotyping and RNA migration patterns could be determined by means of molecular methods like reverse transcriptionpolymerase chain reaction and polyacrylamide gel electrophoresis (PAGE), respectively [11]. However, antigen detection, using EIA or LA, with more than 90% specificity and sensitivity is frequently used for laboratory diagnosis of rotavirus infection [8] .…”

Section: Introductionmentioning

confidence: 99%

Comparison of enzyme immunoassay, latex agglutination and polyacrylamide gel electrophoresis for diagnosis of rotavirus in children

El-Ageery

Ali

El-Khier

et al. 2020

Egyptian Journal of Basic and Applied Sciences

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Rotavirus is considered an important pathogen that causes diarrhea in children. Its laboratory identification is critical for early management and prevention of the outbreak. The aim of this study was to compare the three assays designed for diagnosis of rotavirus in stool samples: polyacrylamide gel electrophoresis (PAGE) as gold standard method, with the antigen detection method using enzyme immunoassay (EIA) and latex agglutination (LA). A cross-sectional retrospective study included 200 stool samples collected from children aged over 5 years with acute gastroenteritis. PAGE, EIA and LA techniques were used for the detection of rotavirus. LA showed higher percentage (36.5%), while EIA and PAGE were nearly equal (32% and 31.5%, respectively) in detection of rotavirus. Compared to PAGE, EIA presented 92.06%, 96.35%, 92.18 and 97.05 of sensitivity, specificity, positive and negative predictive values, respectively. The sensitivity, specificity, positive and negative predictive values of LA test were 84.13%, 85.40%, 72.60% and 92.12%, respectively. Antigen detection using EIA is sensitive, specific and rapid technique for detection of rotavirus infection in stool, while LA may be useful in disease outbreak, or with shortage of means for rotavirus recognition, since it is practical, fast, and reasonably priced.

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RNA-PAGE-Based Diagnosis of Viral Diseases

Kumar,

Kaur,

Sircar

et al. 2022

Springer Protocols Handbooks

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Comparison of agarose gel electrophoresis and RNA-PAGE for rapid detection of rotavirus from faecal samples

Cited by 4 publications

References 19 publications

Molecular Characterization and Risk Factors Associated with Rotavirus among Children with diarrhoea.

Molecular Characterization and Risk Factors Associated with Rotavirus among Children with diarrhoea.

Comparison of enzyme immunoassay, latex agglutination and polyacrylamide gel electrophoresis for diagnosis of rotavirus in children

RNA-PAGE-Based Diagnosis of Viral Diseases

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