Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens.
Vera Gouvea,Roger I. Glass,P. A. Woods,Koki Taniguchi,H. F. Clark,Barbara Forrester,Zhao-Yin Fang +6 more
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TLDR
A PCR typing method was devised in which each human serotype virus produced a characteristic segment size, readily identifiable in agarose gels, which provided a rapid and efficient means of obtaining large quantities of cDNA suitable for sequencing, cloning, and other genetic studies, precluding the need for cell culture and virus purification.Abstract:
The rotavirus gene segment coding for the major outer capsid glycoprotein vp7 was amplified directly from stool specimens by the polymerase chain reaction (PCR). Double-stranded RNA extracted from stool samples was used as the template for reverse transcription, which was followed immediately and in the same reaction mix with amplification, using the Taq polymerase. Various conditions were examined to optimize the yield of the amplified gene. The concentrations of MgCl2, dimethyl sulfoxide, and template RNA were critical. The choice of primer pairs allowed amplification of the entire segment or specific portions. By using type-specific primers derived from distinct regions on the gene, we devised a PCR typing method in which each human serotype virus produced a characteristic segment size, readily identifiable in agarose gels. The PCR typing method was applied to 10 rotavirus reference strains, including all 6 known human serotypes (serotypes 1, 2, 3, 4, 8, and 9), and to 34 stool specimens previously serotyped by an enzyme immunoassay with monoclonal antibodies. An absolute correlation was found between the molecular and serologic methods. In addition, 14 stool specimens nonserotypable by an enzyme immunoassay with monoclonal antibodies could be typed by the PCR method. Besides the application for rotavirus detection and typing directly from stools, the PCR method provides a rapid and efficient means of obtaining large quantities of cDNA suitable for sequencing, cloning, and other genetic studies, precluding the need for cell culture and virus purification.read more
Citations
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Journal ArticleDOI
Evolution of DS-1-like G1P[8] double-gene reassortant rotavirus A strains causing gastroenteritis in children in Vietnam in 2012/2013
Toyoko Nakagomi,Minh Quang Nguyen,Punita Gauchan,Chantal Ama Agbemabiese,Miho Kaneko,Loan Phuong Do,Thiem Dinh Vu,Osamu Nakagomi +7 more
TL;DR: Clinical and phylogenetic analysis suggested that Vietnamese G1P[8] double-gene reassortant strains originated from a locally circulating G2P[4] strain and caused severe diarrhoea, but there was no evidence of increased virulence.
Journal ArticleDOI
Evaluation of circulating intestinally committed memory B cells in children vaccinated with attenuated human rotavirus vaccine.
Olga Lucía Rojas,Liliana Caicedo,Carolina Guzmán,Luz-Stella Rodríguez,Javier Castañeda,Liliana Uribe,Yohanna Andrade,Ricardo Pinzón,Carlos F. Narváez,Juan Manuel Lozano,Beatrice De Vos,Manuel A. Franco,Juana Angel +12 more
TL;DR: When vaccinees and placebo recipients were considered together, a correlation was found between protection from disease and plasma RV IgA measured after dose 2 and RV memory (IgD- CD27+ alpha4beta7+ CCR9+) circulating B cells measured after doses 1, suggesting that other factors are important in explainingprotection from disease.
Journal ArticleDOI
Genomic characterization of human rotavirus G10 strains from the African Rotavirus Network: relationship to animal rotaviruses.
Mathew D. Esona,Krisztián Bányai,Krisztián Bányai,Kimberly Foytich,Molly M. Freeman,Slavica Mijatovic-Rustempasic,Jennifer J. Hull,Tara Kerin,A. D. Steele,George Armah,A. Geyer,Nicola Page,V.A. Agbaya,J.C. Forbi,M. Aminu,Rashi Gautam,L.M. Seheri,James Nyangao,Roger I. Glass,Michael D. Bowen,Jon R. Gentsch +20 more
TL;DR: Although these findings help to elucidate the evolution of ARN G10 strains, additional sequence studies of cognate animal rotavirus genes are needed to determine irrefutably the specific origin of those genes relative to both human and animalRotavirus strains.
Journal ArticleDOI
Molecular epidemiology of rotavirus diarrhoea among children in Haiphong, Vietnam: the emergence of G3 rotavirus.
Ngo Tuan Cuong,Nguyen Binh Minh,Dang Duc Anh,Nguyen Hoai Thu,Nguyen Tuan Tu,Tran Van Nam,Vu Thi Thuy,Michiko Ogino,Md. Mahbub Alam,Toyoko Nakagomi,Osamu Nakagomi,Tetsu Yamashiro +11 more
TL;DR: From September 2006- October 2007 hospital-based surveillance was conducted in Haiphong, Vietnam to determine the distribution of G and P types and electropherotypes of rotavirus, and the emergence of G3P was identified and the strain was predominant among rotaviruses detected.
Journal ArticleDOI
Surveillance Study (2000 to 2001) of G- and P-Type Human Rotaviruses Circulating in South Korea
Bok Soon Min,Yoon Ju Noh,Jinho Shin,Sun Young Baek,Jae Ok Kim,Kyung Il Min,Seung Rel Ryu,Byoug Guk Kim,Do Keun Kim,Seok-Ho Lee,Hong Ki Min,Byung Yoon Ahn,Sue Nie Park +12 more
TL;DR: The commonest G- and P-type combination found in this study was G2P[4], rather than G1P[8], the most prevalent type known worldwide.
References
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Book ChapterDOI
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
Kary B. Mullis,Fred A. Faloona +1 more
TL;DR: A method whereby a nucleic acid sequence can be exponentially amplified in vitro is described in the chapter, and the possibility of utilizing a heat-stable DNA polymerase is explored so as to avoid the need for addition of new enzyme after each cycle of thermal denaturation.
Journal ArticleDOI
Diagnosis of chronic myeloid and acute lymphocytic leukemias by detection of leukemia-specific mRNA sequences amplified in vitro.
Ernest S. Kawasaki,Steven S. Clark,Mazie Y. Coyne,Stephen D. Smith,Richard E. Champlin,Owen N. Witte,Frank McCormick +6 more
TL;DR: D diagnosis of chronic myeloid and acute lymphocytic leukemias by this procedure is rapid, much more sensitive than existing protocols, and independent of the presence or absence of an identifiable Philadelphia chromosome.
Journal ArticleDOI
Serotypic Similarity and Diversity of Rotaviruses of Mammalian and Avian Origin as Studied by Plaque-Reduction Neutralization
TL;DR: A surprising observation that emerged from this study was the existence of a rotavirus (porcine strain SB-1A) bridging serotypes 4 and 5, which was identified in both humans and pigs.
Journal ArticleDOI
Production and preliminary characterization of monoclonal antibodies directed at two surface proteins of rhesus rotavirus.
Harry B. Greenberg,J Valdesuso,K L van Wyke,Karen Midthun,M Walsh,V McAuliffe,R G Wyatt,Anthony R. Kalica,Jorge Flores,Y Hoshino +9 more
TL;DR: Thirty-six monoclonal antibodies immunoprecipitated the 82-kilodalton outer capsid protein, the product of the fourth gene, the viral hemagglutinin, and neutralized rhesus rotavirus to moderate or high titer.
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