title: Development of a new method for diagnosis of Group B Coxsackie genome by reverse transcription loop-mediated isothermal amplification.
abstract: BACKGROUND
  Coxsackie B viruses (genus, Enterovirus; family, Picornaviridae) can cause aseptic meningitis, encephalitis, pleurodynia, and fatal myocarditis, and are implicated in the pathogenesis of dilated cardiomyopathy. The differentiation of the group B Coxsackieviruses into their subtypes has potential clinical and epidemiological implications.
  
  
  OBJECTIVE
  In this study, we developed a one-step, single-tube genogroup-specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of group B Coxsackie genomes targeting 5' UTR region.
  
  
  METHODS
  The amplification can be obtained in less than 1 hour by incubating all the reagents in a single tube with reverse transcriptase and Bst DNA polymerase at 63°C. Detection of gene amplification could be accomplished by agarose gel electrophoresis and the monitoring of gene amplification can also be visualised with the naked eye by using SYBR green I fluorescent dye.
  
  
  RESULTS
  A total of 40 samples comprising 31 positive samples and 9 negative samples were used in this study for comparative evaluation. The results were compared with those from Real-Time Polymerase Chain Reaction (RT-PCR). None of the RT-PCR-positive samples were missed by RT-LAMP, thereby indicating a higher sensitivity of the RT-LAMP assay.
  
  
  CONCLUSIONS
  Thus, due to easy operation without a requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay reported here is extremely rapid, cost-effective, highly sensitive, and specific and has potential usefulness for rapid detection of non-polio enterovirus (NPEV) not only by well-equipped laboratories but also by peripheral diagnostic laboratories with limited financial resources in developing countries.
authors.affiliation: Department of Virology, King Institute of Preventive Medicine, Guindy, Chennai 600 032, India. jaianand.k@lifecellinternational.com
authors.forename: K
authors.initials: K
authors.lastname: Jaianand
authors.forename: N
authors.initials: N
authors.lastname: Saravanan
authors.forename: P
authors.initials: P
authors.lastname: Gunasekaran
authors.forename: A K
authors.initials: AK
authors.lastname: Sheriff
mesh_terms: D052160:Benzothiazoles; D019411:Clinical Laboratory Techniques / Q000191:economics / Q000379:methods*; D003384:Coxsackievirus Infections / Q000175:diagnosis* / Q000821:virology; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I