Differences in Illness Severity among Circulating Norovirus Genotypes in a Large Pediatric Cohort with Acute Gastroenteritis

Norovirus is a major pathogen identified in children with acute gastroenteritis (AGE), little is known about the strain’s diversity and their clinical severity. Stool and/or rectal swabs were collected from children ≤18 years of age recruited at emergency departments (ED), and a provincial nursing advice phone line due to AGE symptoms in the province of Alberta, Canada between December 2014 and August 2018. Specimens were tested using a reverse transcription real time PCR and genotyped by Sanger sequencing. The Modified Vesikari Scale score (MVS) was used to evaluate the disease severity. The objectives are to identify the Genogroup and Genotype distribution and to compare illness severity between the GI and GII genogroups and to complete further analyses comparing the GII genotypes identified. GII.4 was the genotype most commonly identified. Children with GII.4 had higher MVS scores (12.0 (10.0, 14.0; p = 0.002)) and more prolonged diarrheal (5 days (3.0, 7.8)) and vomiting (3.2 days (1.7, 5.3; p < 0.001)) durations compared to other non GII.4 strains. The predominant strain varied by year with GII.4 Sydney[P31] predominant in 2014/15, GII.4 Sydney[P16] in 2015/16 and 2017/18, and GII.3[P12] in 2016/17. Genogroup II norovirus strains predominated in children with AGE with variance between years; clinical severity associated with different strains varied with episodes being most severe among GII.4 infected children.

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Comparison of Common Enrichment Broths Used in Diagnostic Laboratories for Shiga Toxin—Producing Escherichia coli

Microorganisms ◽

10.3390/microorganisms9030503 ◽

2021 ◽

Vol 9 (3) ◽

pp. 503

Author(s):

Michael Bording-Jorgensen ◽

Hannah Tyrrell ◽

Colin Lloyd ◽

Linda Chui

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Real Time Pcr ◽

Acute Gastroenteritis ◽

Diagnostic Testing ◽

First Line ◽

Gram Negative ◽

Culture Independent ◽

Surveillance Programs ◽

Selection Of

Acute gastroenteritis caused by Shiga toxin-producing Escherichia coli (STEC) affects more than 4 million individuals in Canada. Diagnostic laboratories are shifting towards culture-independent diagnostic testing; however, recovery of STEC remains an important aspect of surveillance programs. The objective of this study was to compare common broth media used for the enrichment of STEC. Clinical isolates including O157:H7 as well as non-O157 serotypes were cultured in tryptic soy (TSB), MacConkey (Mac), and Gram-negative (GN) broths and growth was compared using culture on sheep’s blood agar and real-time PCR (qPCR). In addition, a selection of the same isolates was spiked into negative stool and enriched in the same three broths, which were then evaluated using culture on CHROMagarTM STEC agar and qPCR. TSB was found to provide the optimal enrichment for growth of isolates with and without stool. The results from this study suggest that diagnostic laboratories may benefit from enriching STEC samples in TSB as a first line enrichment instead of GN or Mac.

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Development and validation of a reverse transcription real‐time PCR assay for specific detection of PRRSGard vaccine‐like virus

Transboundary and Emerging Diseases ◽

10.1111/tbed.14084 ◽

2021 ◽

Author(s):

Gaurav Rawal ◽

Wannarat Yim‐im ◽

Fabian Chamba ◽

Chad Smith ◽

Jeff Okones ◽

...

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Real Time Pcr ◽

Specific Detection ◽

Pcr Assay ◽

Development And Validation

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Development and application of a novel reverse transcription real-time PCR method for miR-499 quantification

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2013.06.024 ◽

2013 ◽

Vol 46 (15) ◽

pp. 1566-1571 ◽

Cited By ~ 10

Author(s):

Weidong Zheng ◽

Yuwei Di ◽

Yinghong Liu ◽

Ge Huang ◽

Youwei Zheng ◽

...

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Real Time Pcr ◽

Pcr Method

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Seasonal pattern and genotype distribution of sapovirus infection in Japan, 2003–2009

Epidemiology and Infection ◽

10.1017/s0950268811000240 ◽

2011 ◽

Vol 140 (1) ◽

pp. 74-77 ◽

Cited By ~ 20

Author(s):

S. K. DEY ◽

O. PHATHAMMAVONG ◽

T. D. NGUYEN ◽

A. THONGPRACHUM ◽

W. CHAN-IT ◽

...

Keyword(s):

Seasonal Pattern ◽

Age Groups ◽

Cold Season ◽

Viral Gastroenteritis ◽

Genotype Distribution ◽

Chain Reaction ◽

The Family ◽

Causative Agents ◽

Polymerase Chain ◽

Predominant Strain

SUMMARYSapovirus, a member of the family Caliciviridae, is one of the major causative agents of viral gastroenteritis affecting all age groups. A total of 3232 faecal specimens collected from infants and children with gastroenteritis in five different regions of Japan during 2003–2009 were examined for sapovirus by reverse transcription–polymerase chain reaction. Sapoviruses were detected in 131 (4·05%) patients with the peak observed mainly in the cold season (November–March) in Japan during 2003–2009. During the last 6 years, sapovirus GI/1 was the predominant strain in Japan followed by GIV, GII/3, GII/6, GII/2, GII/12 and GI, respectively.

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Analysing CYP2D6*4 Allele frequency in Patients with Schizophrenia

European Psychiatry ◽

10.1016/j.eurpsy.2017.01.314 ◽

2017 ◽

Vol 41 (S1) ◽

pp. S101-S102

Author(s):

V. Djordjevic ◽

T. Jevtovic Stoimenov

Keyword(s):

Allele Frequency ◽

Plasma Concentrations ◽

Clinical Severity ◽

Genotype Distribution ◽

Healthy Individuals ◽

Wild Type ◽

Specific Pcr ◽

Significant Difference ◽

Schizophrenic Patients ◽

The Right

IntroductionSchizophrenia is treated with antipsychotics and other psychotropic medications, many of which are substrates for the highly polymorphic CYP2D6 enzyme. The most frequent variant allele is CYP2D6*4- leading cause of poor metabolism (PM) phenotype. PM causes the reduction of therapeutic response, increase the risk of adverse drug reactions and increase the plasma concentration of both drug and its metabolites above the levels of toxicity.The AimAnalysing CYP2D6*4 allele frequency among schizophrenic patients for further individualisation and rationalisation of therapy.Patients and methodsResearch was conducted on 38 schizophrenic patients and 110 healthy individuals. CYP2D6*4 allele was detected with allele specific PCR.ResultsBoth wild type allele carriers are 55% of the schizophrenic patients, 45% are wild type/*4heterozygous, and *4/*4 homozygous are not identified. There is a statistically significant difference in the genotype distribution (P < 0.05) between schizophrenic patients and healthy individuals. Significantly higher *4 allele frequency (37%) comparing to healthy individuals (P < 0.0001) indicates the necessary caution in administration of CYP2D6 substrates. A lower frequency of PMs in schizophrenic patients than in healthy individuals could be explained with CYP2D6 neuroactive substrate metabolism. Forty-five percent of the schizophrenic patients are intermediate metabolisers carrying the higher risk of adverse response to CYP2D6 substrates comparing to wild type homozygous. As none of the analyzed patients was PM, exceeded plasma concentrations of medications above toxic levels are not expected when administrating the right dosage.ConclusionAltered CYP2D6 metabolism may contribute to the vulnerability, clinical severity and treatment outcome of schizophrenia.Disclosure of interestThe authors have not supplied their declaration of competing interest.

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A stem–loop-mediated reverse transcription real-time PCR for the selective detection and quantification of the replicative strand of an RNA virus

Analytical Biochemistry ◽

10.1016/j.ab.2006.01.046 ◽

2006 ◽

Vol 352 (1) ◽

pp. 120-128 ◽

Cited By ~ 13

Author(s):

Azlinda Anwar ◽

J. Thomas August ◽

Heng Phon Too

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Real Time Pcr ◽

Rna Virus ◽

Selective Detection ◽

Stem Loop ◽

Detection And Quantification

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Detection, Quantitation, and Phylogenetic Analysis of Noroviruses in Japanese Oysters

Applied and Environmental Microbiology ◽

10.1128/aem.69.10.5782-5786.2003 ◽

2003 ◽

Vol 69 (10) ◽

pp. 5782-5786 ◽

Cited By ~ 122

Author(s):

Tomoko Nishida ◽

Hirokazu Kimura ◽

Mika Saitoh ◽

Michiyo Shinohara ◽

Masahiko Kato ◽

...

Keyword(s):

Genetic Diversity ◽

Phylogenetic Analysis ◽

Virus Type ◽

Reverse Transcription ◽

Real Time Pcr ◽

The Other ◽

Capsid Gene ◽

Neighbor Joining ◽

Norwalk Virus ◽

Reverse Transcription Pcr

ABSTRACT Noroviruses (NVs) cause many cases of oyster- or clam-associated gastroenteritis in various countries. We collected 191 samples from Japanese oysters intended for raw consumption that had been harvested from the sea in two different areas between December 2001 and February 2002. To detect, quantitate, and phylogenetically analyze the NV genome in purified concentrates from the stomachs and digestive diverticula of these oysters, we amplified the NV capsid gene by reverse transcription-PCR. Phylogenetic analysis was performed by using the neighbor-joining method. We detected the NV genome in 17 of 191 oysters (9%). Phylogenetic analysis indicated genogroup I (Norwalk virus type) in 3 of the 17 oysters and genogroup II (Snow Mountain virus type) in the other 14. Both genogroups showed wide genetic diversity. To quantify the NV capsid gene in these oysters, we performed real-time PCR using genogroup-specific probes. More than 102 copies of the NV genome were detected in 11 of 17 oysters. The results suggested that about 10% of Japanese oysters intended for raw consumption harbored NVs, and more than 50% of those oysters in which NVs were detected had a large amount.

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