title: Detection of Clostridium perfringens alpha toxin gene in lambs by loop mediated isothermal amplification.
abstract: OBJECTIVE
  The loop mediated isothermal amplification (LAMP) was standardized for rapid detection of Clostridium perfringens.
  
  
  METHODS
  A total of 120 fecal samples were collected from enterotoxemia suspected lambs were used for screening of C. perfringens cpa gene by LAMP. The specificity of the LAMP amplified products was tested by digesting with restriction enzyme XmnI for alpha toxin gene.
  
  
  RESULTS
  Out of 120 samples screened 112 (93.3%) samples were positive by both LAMP and polymerase chain reaction (PCR) for detection of cpa gene which indicated the equal sensitivity of both the tests. The enzyme produced single cut in 162 base pair amplified product of alpha toxin gene at 81 base pair resulting in a single band in gel electrophoresis.
  
  
  CONCLUSIONS
  Both LAMP and PCR for detection of cpa gene indicated the equal sensitivity of both the tests. Standardization of LAMP reaction for amplification of epsilon and beta toxin genes will help to identify the C. perfringens toxin types from the clinical samples. The test could be a suitable alternative to the PCR in detection of toxin types without the help of sophisticated machinery like thermal cycler. Considering its simplicity in operation and high sensitivity, there is the potential use of this technique in clinical diagnosis and surveillance of infectious diseases.
authors.affiliation: State Level Diagnostic Laboratory, College of Veterinary Science, Tirupathi, Andhra Pradesh, India.
authors.forename: B
authors.initials: B
authors.lastname: Radhika
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science, Tirupathi, Andhra Pradesh, India.
authors.forename: N Vinod
authors.initials: NV
authors.lastname: Kumar
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science, Tirupathi, Andhra Pradesh, India.
authors.forename: D
authors.initials: D
authors.lastname: Sreenivasulu
keywords: Clostridium perfringens; enterotoxemia; lambs; loop mediated isothermal amplification