Polymerase Chain Reaction Detection and Typing of Rotaviruses in Fecal Specimens

Gouvêa, Vera; Glass, Roger I.

doi:10.1007/978-3-642-84766-0_26

Cited by 354 publications

(588 citation statements)

References 20 publications

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“…The Rotavirus RT-PCR used the primer sets Beg9/End9 [15] and Con2/Con3 [16], The astrovirus RT-PCR used the primers Mon 244 and Mon 245 [17] and the adenovirus PCR used the primers Hex1DEG/Hex2DEG [18]. The norovirus and sapovirus RT-PCR used primer sets in separate reactions.…”

Section: Laboratory Methodsmentioning

confidence: 99%

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Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Lorrot

Bon

Hajje

et al. 2010

Eur J Clin Microbiol Infect Dis

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International audienceRotavirus is recognised as the most important agent of severe acute gastroenteritis (AGE) in young children. In a 2-year prospective survey, we investigated the epidemiology and clinical features of the viral and bacterial pathogens in children hospitalised for AGE. The study was performed in a Parisian teaching hospital from November 2001 to May 2004. Clinical data were prospectively collected to assess the gastroenteritis severity (20-point Vesikari severity score, the need for intravenous rehydration, duration of hospitalisation). Stools were systematically tested for group A rotavirus, norovirus, astrovirus and adenovirus 40/41, sapovirus and Aichi virus and enteropathogenic bacteria. A total of 457 children (mean age 15.9 months) were enrolled. Viruses were detected in 305 cases (66.7%) and bacteria in 31 cases (6.8%). Rotaviruses were the most frequent pathogen (48.8%), followed by noroviruses (8.3%) and adenoviruses, astroviruses, Aichi viruses and sapoviruses in 3.5%, 1.5%, 0.9% and 0.4%, respectively. Cases of rotavirus gastroenteritis were significantly more severe than those of norovirus with respect to the Vesikari score, duration of hospitalisation and the need for intravenous rehydration. Rotaviruses were the most frequent and most severe cause in children hospitalised for AGE, and noroviruses also account for a large number of cases in this population

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Section: Laboratory Methodsmentioning

confidence: 99%

“…The sex ratio was 1.26. The mean Vesikari score was 11.7 (standard deviation, 3.16; range, [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19]. Among the 95 sets of data rejected in this study, 13 were rejected for the lack of clinical data and 82 for the absence of a faecal sample.…”

Section: Study Population and General Screeningmentioning

confidence: 99%

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Lorrot

Bon

Hajje

et al. 2010

Eur J Clin Microbiol Infect Dis

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“…Rotavirus-positive specimens were categorised into strain G genotype by RT-PCR, as described by Gouvea et al [11]. Rotavirus dsRNA was extracted from stool specimens by standard phenol-chloroform extraction method [11].…”

Section: Rt-pcrmentioning

confidence: 99%

“…Rotavirus dsRNA was extracted from stool specimens by standard phenol-chloroform extraction method [11]. The extracted dsRNA was used in G typing by type-specific primers for RT-PCR.…”

Section: Rt-pcrmentioning

confidence: 99%

Prevalence and Molecular Genotyping of Group A Rotaviruses in Iranian Children

Kargar

Akbarizadeh

2012

Indian J. Virol.

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Rotavirus is the leading cause of acute gastroenteritis in worldwide young children. Effective vaccines to prevent rotavirus infection are currently available, although their clinical use is still limited, and rotavirus still causes many episodes of infantile gastroenteritis, mainly during the winter season. The aim of this study was to evaluate the prevalence of rotavirus infection in children aged \5-years-old who were hospitalised for gastroenteritis. One hundred and sixty-three stool samples from hospitalised children (\5-years-old) complicated with severe diarrhoea, in two hospitals in Jahrom City, Iran were collected from 2009 to 2010. Antigenic prevalence of rotavirus group A was distinguished by enzyme immunoassay. The antigen of group A rotavirus was diagnosed by EIA in 75 of 163 collected samples. The genotype of EIA-positive samples was determined by nested RT-PCR. The frequency of rotavirus genotypes G1, G2, G3, G4 and G9 was 17.33, 13.34, 2.67, 30.66 and 2.67 %, respectively. Also, the frequency of mixed and non-typable genotypes was detected in 2.67 and 30.66 %, respectively. G1/G8 mixed infection was the first of these rotavirus genotypes to be reported in Iran. Detection of high prevalence of group A rotavirus infection in hospitalised children with diarrhoea, and determination of circulating rotavirus genotypes in this region of Iran, provide useful data for formulating effective vaccines; especially for infants less than 5-years-old.

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“…Positive and negative controls were also included. One-step semi-nested multiplex reverse transcriptase polymerase chain reaction (RT-PCR) using SuperScript One-Step RT-PCR with Platinum ® Taq (Invitrogen, USA) and one set of primers described by Das , a random primed reverse transcription step followed by a two-step PCR procedure was implemented where the consensus primers VP7-F/VP7-R and VP4-F/VP4-R were used for the first PCR reactions for G and P typing, respectively, while the second PCR assays were performed by using mixes of one consensus and type-specific primers as follows: for G genotype determination (VP7-R and aBT1/ G1, aCT2/G2, mG3/G3, aDT4/G4, aAT8/G8, mG9/G9, G10, G12) [23,31,32], and for P genotype detection (VP4-F and 1 T-1D, 2 T-1, 3 T-1, 4 T-1, 5 T-1, P [11]) [30,33,34].…”

Section: G and P Genotypingmentioning

confidence: 99%

Molecular epidemiology of rotaviruses in Bulgaria: annual shift of the predominant genotype

Mladenova

Korsun²,

Geonova³

et al. 2010

Eur J Clin Microbiol Infect Dis

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Polymerase Chain Reaction Detection and Typing of Rotaviruses in Fecal Specimens

Cited by 354 publications

References 20 publications

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Epidemiology and clinical features of gastroenteritis in hospitalised children: prospective survey during a 2-year period in a Parisian hospital, France

Prevalence and Molecular Genotyping of Group A Rotaviruses in Iranian Children

Molecular epidemiology of rotaviruses in Bulgaria: annual shift of the predominant genotype

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