Open Access
R esearch
(Published
online:
20-01-2016)
11.
Detection of
Clostridium perfringens
alpha toxin gene in lambs by loop mediated
isothermal amplification -
B. Radhika, N. Vinod Kumar and D. Sreenivasulu
Veterinary World, 9(1): 60-64
doi:
10.14202/vetworld.2016.60-64
B. Radhika:
State Level Diagnostic Laboratory, College of Veterinary
Science, Tirupathi, Andhra Pradesh, India; radhi.radhi.71@gmail.com
N. Vinod Kumar:
Department of Veterinary Microbiology, College of Veterinary
Science, Tirupathi, Andhra Pradesh, India; nagaram_vinod@yahoo.com
D. Sreenivasulu:
Department of Veterinary Microbiology, College of Veterinary
Science, Tirupathi, Andhra Pradesh, India; dsreenivasulu10@gmail.com
Received: 20-08-2015, Revised: 02-12-2015, Accepted: 11-12-2015,
Published online: 20-01-2016
Corresponding author:
B. Radhika, e-mail: radhi.radhi.71@gmail.com
Citation:
Radhika B, Kumar NV, Sreenivasulu D (2016) Detection of
Clostridium perfringens
alpha toxin gene in lambs by loop mediated isothermal
amplification,
Veterinary World
9(1): 60-64.
Abstract
Aim:
The loop mediated isothermal amplification (LAMP) was
standardized for rapid detection of
Clostridium perfringens.
Materials and 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.
Conclusion:
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.
Keywords:
Clostridium perfringens,
enterotoxemia, lambs, loop mediated isothermal amplification.
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