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Marize Pereira Miagostovich - One of the best experts on this subject based on the ideXlab platform.
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RotAvirus A shedding And hbgA host genetic susceptibility in A birth community cohort rio de jAneiro brAzil 2014 2018
Scientific Reports, 2020Co-Authors: Carina Pacheco Cantelli, Denise Cotrim Da Cunha, Patricia Brasil, Johan Nordgren, Lennart Svensson, Alvaro Jorge Velloso, Rosane Maria Santos De Assis, Jose Junior Franca De Barros, Francisco C A Mello, Marize Pereira MiagostovichAbstract:Recent studies hAve investigAted whether the humAn histo-blood group Antigen (HBGAs) could Affect the effectiveness of the orAl RotAvirus vAccines, suggesting secretor positive individuAls develop A more robust response. We investigAted the RotAvirus A (RVA) shedding in AssociAtion with the host susceptibility profile in children from A birth community-cohort in Rio de JAneiro, BrAzil, from 2014 to 2018. A totAl of 132 children were followed-up between 0 to 11-month-old, stool sAmples were collected before/After the 1st/2nd RV1 vAccinAtion doses And sAlivA sAmples were collected during the study. RVA shedding wAs screened by RT-qPCR And G/P genotypes determined by multiplex RT-PCR And/or SAnger nucleotide sequencing. The sequencing indicAted An F167L Amino Acid chAnge in the RV1 VP8* P[8] in 20.5% of shedding follow-ups And these mutAnt subpopulAtions were quAntified by pyrosequencing. The HBGA/secretor stAtus wAs determined And 80.3% of the children were secretors. Twenty-one FUT2 gene SNPs were identified And two new mutAtions were observed. The mutAnt F167L RV1 VP8* P[8] wAs detected significAntly more in Le (A+b+) secretors (90.5%) compAred to non-secretors And even to secretors Le (A−b+) (9.5%). The study highlights the probAble AssociAtion between RV1 shedding And HBGAs As A mArker for evAluAting vAccine strAin host susceptibility.
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norovirus infection And hbgA host genetic susceptibility in A birth community cohort rio de jAneiro brAzil
Infection Genetics and Evolution, 2020Co-Authors: Carina Pacheco Cantelli, Fabio Correia Malta, Denise Cotrim Da Cunha, Patricia Brasil, Marize Pereira Miagostovich, Marcia Terezinha Baroni De Moraes, Tulio Machado Fumian, Johan Nordgren, Lennart Svensson, Jose Paulo Gagliardi LeiteAbstract:Norovirus hAs emerged As An importAnt virAl Agent of Acute pediAtric gAstroenteritis, with A growing genetic diversity reported in the lAst decAdes. Histo-blood group Antigens (HBGAs) present on the surfAce of enterocytes Are susceptibility fActors for norovirus infection And differ between populAtions which could Affects the epidemiology And evolution of these viruses. This study investigAted the frequency, incidence And genetic diversity of noroviruses in A cohort of RotAvirus A vAccinAted children in AssociAtion to the host HBGA (Secretor/Lewis) genetic susceptibility profile. Norovirus genogroups I And II (GI/GII) were screened by RT-qPCR in 569 stool sAmples from 132 children followed-up from birth to 11 months of Age during 2014--2018. Noroviruses were identified in 21.2% of children enrolled in this study, with A norovirus detection rAte of 5.6% (32/569), in 17.1% And 4.7% of Acute diArrheic episodes (ADE) And non-ADE, respectively. The norovirus incidence wAs 5.8 infections per 100 child-months. PArtiAl nucleotide sequencing chArActerized six different norovirus genotypes, with GII.4 Sydney 2012 being detected in 50% AssociAted with three different polymerAse genotypes (GII·P31, GII·P16 And GII·P4 New OrleAns 2009). FUT3 genotyping wAs yielded seven new mutAtions in this populAtion. A significAnt AssociAtion between symptomAtic norovirus infection And secretor profile could be inferred.
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GAstroenteric Viruses Detection in A Drinking WAter Distribution-to-Consumption System in A Low-Income Community in Rio de JAneiro
Food and Environmental Virology, 2020Co-Authors: Marize Pereira Miagostovich, Fabio Correia Malta, Mônica Simões Rocha, Fabiane Bertoni Reis, Marcelo Santos Sampaio, Renata Saldanha Da Gama Gracie Carrijo, Janaína Rodrigues, Amanda Genuino, Matheus Ribeiro Da Silva Assis, Tulio Machado FumianAbstract:The AvAilAbility of drinking wAter is one of the mAin determinAnts of quAlity of life, diseAse prevention And the promotion of heAlth. Viruses Are importAnt Agents of wAterborne diseAses And hAve been described As importAnt mArkers of humAn fAecAl contAminAtion. This study Aimed to investigAte viruses’ presence As An indicAtor of drinking wAter quAlity in low-income communities in the MAnguinhos AreA, Rio de JAneiro, BrAzil. Three hundred And four drinking wAter sAmples (2L/eAch) were collected Along the drinking wAter distribution-to-consumption pAthwAy in households, As well As heAlthcAre And school units. WAter sAmples were collected both directly from the wAter supply prior to distribution And After storAge in tAnks And filtrAtion units. Using qPCR, viruses were detected 50 times in 45 wAter sAmples (15%), 19 of these being humAn Adenovirus, 17 RotAvirus A And 14 norovirus GII. VirAl loAds recovered rAnged from 5E+10 to 8.7E+10^6 genome copies/Liter. Co-detection wAs observed in five household wAter sAmples And there wAs no difference regArding virus detection Across sAmpling sites. PrecArious And inAdequAte environmentAl conditions chArActerized by the lAck of locAl infrAstructure regArding bAsic sAnitAtion And wAste collection in the territory, As well As negligent hygiene hAbits, could explAin virAl detection in drinking wAter in regions with A wAter supply system.
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SurveillAnce of Enteric Viruses And ThermotolerAnt Coliforms in SurfAce WAter And BivAlves from A MAngrove EstuAry in SoutheAstern BrAzil
Food and Environmental Virology, 2019Co-Authors: Regina Keller, Marize Pereira Miagostovich, Tulio Machado Fumian, Rodrigo Pratte-santos, Karolina Scarpati, Sara Angelino Martins, Suzanne Mariane Loss, Sérvio Túlio CassiniAbstract:This study wAs conducted to evAluAte the microbiologicAl quAlity of A mAngrove estuAry in the VitóriA BAy region, Espírito SAnto, BrAzil. We AnAlyzed the presence And concentrAtion of enteric viruses And thermotolerAnt coliforms in wAter, mussels ( MytellA chArruAnA And MytellA guyAnensis ), And oysters ( CrAssostreA rhizophorAe ), collected over A 13-month period. HumAn Adenovirus, RotAvirus A (RVA), And norovirus genogroup II were AnAlyzed by quAntitAtive PCR. The highest virAl loAd wAs found in RVA-positive sAmples with A concentrAtion of 3.0 × 10^4 genome copies (GC) L^−1 in wAter sAmples And 1.3 × 10^5 GC g^−1 in bivAlves. RVA wAs the most prevAlent virus in All mAtrices. ThermotolerAnt coliforms were quAntified As colony-forming units (CFU) by the membrAne filtrAtion method. The concentrAtion of these bActeriA in wAter wAs in AccordAnce with the BrAziliAn stAndArd for recreAtionAl wAters (
Tulio Machado Fumian - One of the best experts on this subject based on the ideXlab platform.
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norovirus infection And hbgA host genetic susceptibility in A birth community cohort rio de jAneiro brAzil
Infection Genetics and Evolution, 2020Co-Authors: Carina Pacheco Cantelli, Fabio Correia Malta, Denise Cotrim Da Cunha, Patricia Brasil, Marize Pereira Miagostovich, Marcia Terezinha Baroni De Moraes, Tulio Machado Fumian, Johan Nordgren, Lennart Svensson, Jose Paulo Gagliardi LeiteAbstract:Norovirus hAs emerged As An importAnt virAl Agent of Acute pediAtric gAstroenteritis, with A growing genetic diversity reported in the lAst decAdes. Histo-blood group Antigens (HBGAs) present on the surfAce of enterocytes Are susceptibility fActors for norovirus infection And differ between populAtions which could Affects the epidemiology And evolution of these viruses. This study investigAted the frequency, incidence And genetic diversity of noroviruses in A cohort of RotAvirus A vAccinAted children in AssociAtion to the host HBGA (Secretor/Lewis) genetic susceptibility profile. Norovirus genogroups I And II (GI/GII) were screened by RT-qPCR in 569 stool sAmples from 132 children followed-up from birth to 11 months of Age during 2014--2018. Noroviruses were identified in 21.2% of children enrolled in this study, with A norovirus detection rAte of 5.6% (32/569), in 17.1% And 4.7% of Acute diArrheic episodes (ADE) And non-ADE, respectively. The norovirus incidence wAs 5.8 infections per 100 child-months. PArtiAl nucleotide sequencing chArActerized six different norovirus genotypes, with GII.4 Sydney 2012 being detected in 50% AssociAted with three different polymerAse genotypes (GII·P31, GII·P16 And GII·P4 New OrleAns 2009). FUT3 genotyping wAs yielded seven new mutAtions in this populAtion. A significAnt AssociAtion between symptomAtic norovirus infection And secretor profile could be inferred.
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GAstroenteric Viruses Detection in A Drinking WAter Distribution-to-Consumption System in A Low-Income Community in Rio de JAneiro
Food and Environmental Virology, 2020Co-Authors: Marize Pereira Miagostovich, Fabio Correia Malta, Mônica Simões Rocha, Fabiane Bertoni Reis, Marcelo Santos Sampaio, Renata Saldanha Da Gama Gracie Carrijo, Janaína Rodrigues, Amanda Genuino, Matheus Ribeiro Da Silva Assis, Tulio Machado FumianAbstract:The AvAilAbility of drinking wAter is one of the mAin determinAnts of quAlity of life, diseAse prevention And the promotion of heAlth. Viruses Are importAnt Agents of wAterborne diseAses And hAve been described As importAnt mArkers of humAn fAecAl contAminAtion. This study Aimed to investigAte viruses’ presence As An indicAtor of drinking wAter quAlity in low-income communities in the MAnguinhos AreA, Rio de JAneiro, BrAzil. Three hundred And four drinking wAter sAmples (2L/eAch) were collected Along the drinking wAter distribution-to-consumption pAthwAy in households, As well As heAlthcAre And school units. WAter sAmples were collected both directly from the wAter supply prior to distribution And After storAge in tAnks And filtrAtion units. Using qPCR, viruses were detected 50 times in 45 wAter sAmples (15%), 19 of these being humAn Adenovirus, 17 RotAvirus A And 14 norovirus GII. VirAl loAds recovered rAnged from 5E+10 to 8.7E+10^6 genome copies/Liter. Co-detection wAs observed in five household wAter sAmples And there wAs no difference regArding virus detection Across sAmpling sites. PrecArious And inAdequAte environmentAl conditions chArActerized by the lAck of locAl infrAstructure regArding bAsic sAnitAtion And wAste collection in the territory, As well As negligent hygiene hAbits, could explAin virAl detection in drinking wAter in regions with A wAter supply system.
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SurveillAnce of Enteric Viruses And ThermotolerAnt Coliforms in SurfAce WAter And BivAlves from A MAngrove EstuAry in SoutheAstern BrAzil
Food and Environmental Virology, 2019Co-Authors: Regina Keller, Marize Pereira Miagostovich, Tulio Machado Fumian, Rodrigo Pratte-santos, Karolina Scarpati, Sara Angelino Martins, Suzanne Mariane Loss, Sérvio Túlio CassiniAbstract:This study wAs conducted to evAluAte the microbiologicAl quAlity of A mAngrove estuAry in the VitóriA BAy region, Espírito SAnto, BrAzil. We AnAlyzed the presence And concentrAtion of enteric viruses And thermotolerAnt coliforms in wAter, mussels ( MytellA chArruAnA And MytellA guyAnensis ), And oysters ( CrAssostreA rhizophorAe ), collected over A 13-month period. HumAn Adenovirus, RotAvirus A (RVA), And norovirus genogroup II were AnAlyzed by quAntitAtive PCR. The highest virAl loAd wAs found in RVA-positive sAmples with A concentrAtion of 3.0 × 10^4 genome copies (GC) L^−1 in wAter sAmples And 1.3 × 10^5 GC g^−1 in bivAlves. RVA wAs the most prevAlent virus in All mAtrices. ThermotolerAnt coliforms were quAntified As colony-forming units (CFU) by the membrAne filtrAtion method. The concentrAtion of these bActeriA in wAter wAs in AccordAnce with the BrAziliAn stAndArd for recreAtionAl wAters (
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detection And moleculAr chArActerizAtion of cAliciviruses vesivirus And norovirus in An outbreAk of Acute diArrheA in kittens from brAzil
Veterinary Journal, 2015Co-Authors: Tatiana Xavier De Castro, Peter A. White, Rita De Cassia Nasser Cubel Garcia, Tulio Machado Fumian, E M Costa, Renata Mello, Jose Paulo Gagliardi LeiteAbstract:Feline cAliciviruses (FCVs) hAve occAsionAlly been described in cAts in AssociAtion with enteric diseAse, but An etiologicAl role for these viruses in Acute gAstroenteritis is still uncleAr. In this study, moleculAr chArActerizAtion of FCV And feline norovirus (FNoV) wAs undertAken using reAl-time PCR (RT-PCR) And sequence AnAlysis of the ORF1 region in fecAl specimens from 29 diArrheic cAts. The specimens were Also screened for pArvovirus, coronAvirus, Astrovirus And group A RotAvirus. A quAntitAtive one step RT-PCR wAs Also performed to detect And quAntitAte NoV genogroup IV And the role of these AnimAl cAliciviruses in feline gAstroenteritis wAs investigAted. This is the first description of enteric FCV And FNoV in South AmericA.
Takako Utsumi - One of the best experts on this subject based on the ideXlab platform.
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Improvement of RotAvirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set
Frontiers Media S.A., 2019Co-Authors: Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Yoshiki Fujii, Maria Inge Lusida, Ikuo Shoji, Kazuhiko KatayamaAbstract:RotAvirus A (RVA) is A mAjor cAuse of gAstroenteritis in infAnts And young children. After vAccine introduction, RVA surveillAnce hAs become more importAnt for monitoring chAnges in genotype distribution, And the semi-nested multiplex-PCR is A populAr method for RVA genotyping. In pArticulAr, the VP7 primer set reported by GouveA And colleAgues in 1990 is still widely used worldwide As the recommended WHO primer set in regionAl And nAtionAl reference RVA surveillAnce lAborAtories. However, this primer set yielded some mistAkes with recent epidemic strAins. The newly emerged equine-like G3 strAins were mistyped As G1, G8 strAins were mistyped As G3, the G9 lineAge 3 strAins showed very weAk bAnd, And the G9 lineAge 6 strAins showed A G9-specific bAnd And A non-specific bAnd. GouveA’s stAndArd protocol hAs become relAtively unreliAble for identifying genotypes correctly. To overcome this limitAtion, we redesigned the primer set to include recent epidemic strAins. Our new primer set enAbled us to correctly identify the VP7 genotypes of representAtive epidemic strAins by AgArose gel electrophoresis (G1, G2, humAn typicAl G3, equine-like G3, G4, G8, G9, And G12). We believe thAt the multiplex-PCR method with our new primer set is A useful And vAluAble tool for surveillAnce of RVA epidemics
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ImAge_3_Improvement of RotAvirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set.TIF
2019Co-Authors: Yoshiki Fujii, Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Maria Inge Lusida, Ikuo Shoji, Kazuhiko KatayamaAbstract:RotAvirus A (RVA) is A mAjor cAuse of gAstroenteritis in infAnts And young children. After vAccine introduction, RVA surveillAnce hAs become more importAnt for monitoring chAnges in genotype distribution, And the semi-nested multiplex-PCR is A populAr method for RVA genotyping. In pArticulAr, the VP7 primer set reported by GouveA And colleAgues in 1990 is still widely used worldwide As the recommended WHO primer set in regionAl And nAtionAl reference RVA surveillAnce lAborAtories. However, this primer set yielded some mistAkes with recent epidemic strAins. The newly emerged equine-like G3 strAins were mistyped As G1, G8 strAins were mistyped As G3, the G9 lineAge 3 strAins showed very weAk bAnd, And the G9 lineAge 6 strAins showed A G9-specific bAnd And A non-specific bAnd. GouveA’s stAndArd protocol hAs become relAtively unreliAble for identifying genotypes correctly. To overcome this limitAtion, we redesigned the primer set to include recent epidemic strAins. Our new primer set enAbled us to correctly identify the VP7 genotypes of representAtive epidemic strAins by AgArose gel electrophoresis (G1, G2, humAn typicAl G3, equine-like G3, G4, G8, G9, And G12). We believe thAt the multiplex-PCR method with our new primer set is A useful And vAluAble tool for surveillAnce of RVA epidemics.
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equine like g3 RotAvirus strAins As predominAnt strAins Among children in indonesiA in 2015 2016
Infection Genetics and Evolution, 2018Co-Authors: Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Zayyin Dinana, Soegeng Soegijanto, Yoshiki Fujii, Laura Navika Yamani, Chieko Matsui, Lin DengAbstract:AbstrAct RotAvirus A (RVA) is A mAjor cAuse of Acute gAstroenteritis in humAns And AnimAls worldwide. As A result of the segmented nAture of the RotAvirus genome, genetic reAssortment commonly occurs. This study Aims to clArify the genetic chArActeristics of RVAs circulAting in IndonesiA. From June 2015 through August 2016, stool sAmples were collected from 134 children Aged
Yen Hai Doan - One of the best experts on this subject based on the ideXlab platform.
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Improvement of RotAvirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set
Frontiers Media S.A., 2019Co-Authors: Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Yoshiki Fujii, Maria Inge Lusida, Ikuo Shoji, Kazuhiko KatayamaAbstract:RotAvirus A (RVA) is A mAjor cAuse of gAstroenteritis in infAnts And young children. After vAccine introduction, RVA surveillAnce hAs become more importAnt for monitoring chAnges in genotype distribution, And the semi-nested multiplex-PCR is A populAr method for RVA genotyping. In pArticulAr, the VP7 primer set reported by GouveA And colleAgues in 1990 is still widely used worldwide As the recommended WHO primer set in regionAl And nAtionAl reference RVA surveillAnce lAborAtories. However, this primer set yielded some mistAkes with recent epidemic strAins. The newly emerged equine-like G3 strAins were mistyped As G1, G8 strAins were mistyped As G3, the G9 lineAge 3 strAins showed very weAk bAnd, And the G9 lineAge 6 strAins showed A G9-specific bAnd And A non-specific bAnd. GouveA’s stAndArd protocol hAs become relAtively unreliAble for identifying genotypes correctly. To overcome this limitAtion, we redesigned the primer set to include recent epidemic strAins. Our new primer set enAbled us to correctly identify the VP7 genotypes of representAtive epidemic strAins by AgArose gel electrophoresis (G1, G2, humAn typicAl G3, equine-like G3, G4, G8, G9, And G12). We believe thAt the multiplex-PCR method with our new primer set is A useful And vAluAble tool for surveillAnce of RVA epidemics
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ImAge_3_Improvement of RotAvirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set.TIF
2019Co-Authors: Yoshiki Fujii, Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Maria Inge Lusida, Ikuo Shoji, Kazuhiko KatayamaAbstract:RotAvirus A (RVA) is A mAjor cAuse of gAstroenteritis in infAnts And young children. After vAccine introduction, RVA surveillAnce hAs become more importAnt for monitoring chAnges in genotype distribution, And the semi-nested multiplex-PCR is A populAr method for RVA genotyping. In pArticulAr, the VP7 primer set reported by GouveA And colleAgues in 1990 is still widely used worldwide As the recommended WHO primer set in regionAl And nAtionAl reference RVA surveillAnce lAborAtories. However, this primer set yielded some mistAkes with recent epidemic strAins. The newly emerged equine-like G3 strAins were mistyped As G1, G8 strAins were mistyped As G3, the G9 lineAge 3 strAins showed very weAk bAnd, And the G9 lineAge 6 strAins showed A G9-specific bAnd And A non-specific bAnd. GouveA’s stAndArd protocol hAs become relAtively unreliAble for identifying genotypes correctly. To overcome this limitAtion, we redesigned the primer set to include recent epidemic strAins. Our new primer set enAbled us to correctly identify the VP7 genotypes of representAtive epidemic strAins by AgArose gel electrophoresis (G1, G2, humAn typicAl G3, equine-like G3, G4, G8, G9, And G12). We believe thAt the multiplex-PCR method with our new primer set is A useful And vAluAble tool for surveillAnce of RVA epidemics.
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equine like g3 RotAvirus strAins As predominAnt strAins Among children in indonesiA in 2015 2016
Infection Genetics and Evolution, 2018Co-Authors: Takako Utsumi, Rury Mega Wahyuni, Yen Hai Doan, Zayyin Dinana, Soegeng Soegijanto, Yoshiki Fujii, Laura Navika Yamani, Chieko Matsui, Lin DengAbstract:AbstrAct RotAvirus A (RVA) is A mAjor cAuse of Acute gAstroenteritis in humAns And AnimAls worldwide. As A result of the segmented nAture of the RotAvirus genome, genetic reAssortment commonly occurs. This study Aims to clArify the genetic chArActeristics of RVAs circulAting in IndonesiA. From June 2015 through August 2016, stool sAmples were collected from 134 children Aged
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g8 RotAviruses with conserved genotype constellAtions detected in mAlAwi over 10 yeArs 1997 2007 displAy frequent gene reAssortment Among strAins co circulAting in humAns
Journal of General Virology, 2013Co-Authors: Toyoko Nakagomi, Osamu Nakagomi, Miren Iturrizagomara, Yen Hai Doan, Winifred Dove, Bagrey Ngwira, Nigel A CunliffeAbstract:RotAvirus A, the most common cAuse of severe diArrhoeA in children worldwide, occurs in five mAjor VP7 (G) And VP4 (P) genotype combinAtions, comprising G1P[8], G2P[4], G3P[8], G4P[8] And G9P[8]. However, G8, A common bovine RotAvirus genotype, hAs been reported frequently Among children in AfricAn countries. SurveillAnce of RotAvirus gAstroenteritis conducted in A sentinel hospitAl in BlAntyre, MAlAwi between 1997 And 2007 provided A rAre opportunity to exAmine the whole genotype constellAtion of G8 strAins And their evolution over time. A sAmple of 27 (9.0 %) of 299 G8 strAins wAs selected to represent eAch surveillAnce yeAr And A rAnge of P genotypes, which shifted in predominAnce from P[6] to P[4] And P[8] during the study period. Following cell culture AdAptAtion, whole genome sequencing demonstrAted thAt the genetic bAckground of 26 strAins possessed the DS-1 genotype constellAtion. A single G8P[6] strAin wAs A reAssortAnt in which both NSP2 And NSP5 genes from strAins with the WA genotype constellAtion hAd been inserted into A strAin with the DS-1 genotype bAckground. Phylogenetic AnAlysis suggested frequent reAssortment Among co-circulAting strAins with the DS-1 genotype constellAtion. Little evidence wAs identified to suggest the introduction of contemporAry bovine RotAvirus genes into Any of the 27 G8 strAins exAmined. In conclusion, MAlAwiAn G8 strAins Are closely relAted to other humAn strAins with the DS-1 genotype constellAtion. They hAve evolved over the lAst decAde through genetic reAssortment with other humAn RotAviruses, chAnging their VP4 genotypes while mAintAining A conserved genotype constellAtion for the remAining structurAl And non-structurAl proteins.
Jim Gray - One of the best experts on this subject based on the ideXlab platform.
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diAgnosing RotAvirus A AssociAted iid using elisA to identify A cut off for reAl time rt pcr
Journal of Clinical Virology, 2009Co-Authors: Gemma Phillips, Ben Lopman, Clarence C Tam, Miren Iturrizagomara, David W Brown, Jim GrayAbstract:BAckground The use of RT-PCR for diAgnosis of group A RotAviruses is increAsing, but up to 14% of heAlthy individuAls mAy be positive by RT-PCR. If RT-PCR is not well correlAted with diseAse, RotAvirus A mAy not AlwAys be the cAuse of illness in RT-PCR positive pAtients with infectious intestinAl diseAse (IID). Objectives To describe the differences in fAecAl virAl loAd between ELISA positive IID cAses, RT-PCR positive cAses And heAlthy controls. To develop A cut-off in fAecAl virAl loAd for Attributing illness to RotAvirus A in RT-PCR positive IID cAses. Study design FAecAl virAl loAd wAs meAsured, using reAl time RT-PCR, in 118 community IID cAses And 65 heAlthy controls, previously tested by ELISA. Cycle threshold (Ct) vAlues from the reAl-time RT-PCR were used As A proxy meAsure of virAl loAd. A cut-off for Attributing illness to RotAvirus A wAs selected, using ROC AnAlysis. Results There wAs little overlAp in virAl loAd between ELISA positive IID cAses (mediAn Ct 17) And heAlthy controls (mediAn Ct 37), but ELISA negAtive, RT-PCR positive IID cAses hAd virAl loAds similAr to heAlthy controls (mediAn Ct 37), indicAting thAt RT-PCR is not detecting extrA cAses of group A RotAvirus AssociAted IID, only sub-clinicAl infections. The optimAl cut-off in the reAl time RT-PCR wAs At Ct vAlue 24 to 27. Conclusion ELISA is the best method for the lAborAtory diAgnosis of RotAvirus A AssociAted IID. If RT-PCR is used, it is AdvisAble to use A reAl time plAtform And to use A virAl loAd cut-off equivAlent to the detection limit of ELISA.
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detection by pcr of eight groups of enteric pAthogens in 4 627 fAecAl sAmples re exAminAtion of the english cAse control infectious intestinAl diseAse study 1993 1996
European Journal of Clinical Microbiology & Infectious Diseases, 2007Co-Authors: C F L Amar, Miren Iturrizagomara, Jim Gray, C L East, E Maclure, J MclauchlinAbstract:The English cAse-control Infectious IntestinAl DiseAse Study (1993–1996) fAiled to detect An enteric pAthogen or toxin in 49% of cAses of gAstroenteritis. In the present study, polymerAse chAin reAction (PCR) AssAys were Applied to DNA And cDNA generAted from 4,627 fAecAl sAmples from cAses And controls Archived during the originAl study for the detection of norovirus, RotAvirus, sApovirus, CAmpylobActer spp., SAlmonellA spp., enteroAggregAtive EscherichiA coli, Cryptosporidium spp., And GiArdiA spp. The percentAge of Archived sAmples from cAses And from controls in which At leAst one Agent (or toxin) wAs detected increAsed from 53% in the originAl study to 75% And from 19 to 42%, respectively, After the ApplicAtion of PCR AssAys. Among cAses, the following percentAges of enteric pAthogens were detected: norovirus 36%, RotAvirus A 31%, sApovirus 4%, SAlmonellA spp. 6%, CAmpylobActer jejuni 13%, CAmpylobActer coli 2%, other CAmpylobActer spp. 8%, enteroAggregAtive E. coli 6%, GiArdiA spp. 2%, And Cryptosporidium spp. 2%. The present study provides AdditionAl insight into the Aetiology of infectious intestinAl diseAse in EnglAnd And highlights the occurrence of virAl infections in cAses As well As in AsymptomAtic individuAls. Other notAble findings include the frequent presence of CAmpylobActer spp. other thAn C. jejuni or C. coli, the high frequency of multiple Agents in 41% of cAses And in 13% of controls, And the vAriAtion in the Aetiology And rAte of infection found for different Age groups. The results demonstrAte the greAter sensitivity of PCR-bAsed methods compAred to current conventionAl methods.
