Genomic Fingerprinting of the Vaccine Strain of Clostridium Tetani by Restriction Fragment Length Polymorphism Technique

ahmad sakhravi; Esmaile Asli; Naser Harzandi; Mohammad-Mehdi Feizabadi; Abdolreza Movahedi; Nader Mosavari; davoud sadeghi

Zahedan Journal of Research in Medical Sciences

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Genomic Fingerprinting of the Vaccine Strain of Clostridium Tetani by Restriction Fragment Length Polymorphism Technique

Author(s):

ahmad sakhravi^1,*,

Esmaile Asli¹,

Naser Harzandi¹,

Mohammad-Mehdi Feizabadi²,

Abdolreza Movahedi³,

Nader Mosavari³,

davoud sadeghi⁴

1Department of Microbiology, Faculty of Sciences, Islamic Azad University, Karaj Branch, Karaj, Iran

2Department of Microbiology, Tehran University of Medical Sciences, Tehran, Iran

3Department of Microbiology, Razi Vaccine & Serum Research Institute, Tehran University of Medical Sciences, Tehran, Iran

4Master of Biophysics, Razi Vaccine & Serum Research Institute

Zahedan Journal of Research in Medical Sciences:Vol. 15, issue 5; e92974

Published online:Nov 18, 2012

Article type:Research Article

Received:Feb 24, 2012

Accepted:Oct 22, 2012

How to Cite:ahmad sakhraviEsmaile AsliNaser HarzandiMohammad-Mehdi FeizabadiAbdolreza MovahediNader Mosavaridavoud sadeghiet al.Genomic Fingerprinting of the Vaccine Strain of Clostridium Tetani by Restriction Fragment Length Polymorphism Technique.Zahedan J Res Med Sci.15(5):e92974.

Abstract

Background : Clostridium tetani or Nicolaier’s bacillus is an obligatory anaerobic, Gram-positive, movable with terminal or sub terminal spore. The chromosome of C. tetani contains 2,799,250 bp with a G+C content of 28.6%. The aim of this study was identification and genomic fingerprinting of the vaccine strain of C. tetani.
Materials and Methods : The vaccine strain of C. tetani was provided by Razi Vaccine and Serum Research Institute. The seeds were inoculated into Columbia blood agar and grown for 72 h and transferred to the thioglycolate broth medium for further 36 h culturing. The cultures were incubated at 35ºC in anaerobic conditions. DNA extraction with phenol/ chloroform method was performed. After extraction, the consistency of DNA was assayed. Next, the vaccine strain was digested using pvuII enzyme and incubated at 37ºC for overnight. The digested DNA was gel-electrophoresed by 1% agarose for a short time. Then, the gel was studied with Gel Doc system and transferred to Hybond N+membrane using standard DNA blotting techniques.
Results : The vaccine strain of C. tetani genome was fingerprinted by RFLP technique. Our preliminary results showed no divergence exists in the vaccine strain used for the production tetanus toxoid during the periods of 1990-2011.
Conclusion : Observation suggests that there is lack of significant changes in RFLP genomic fingerprinting profile of the vaccine strain. Therefore, this strain did not lose its efficiency in tetanus vaccine production. RFLP analysis is worthwhile in investigating the nature of the vaccine strain C. tetani.

Keywords

Clostridium tetani

Vaccinal strain

RFLP technique

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