Tropical Animal Health and Production

, Volume 51, Issue 6, pp 1559–1569 | Cite as

Molecular detection and characterization of Clostridium perfringens toxin genes causing necrotic enteritis in broiler chickens

  • N. Praveen KumarEmail author
  • N. Vinod Kumar
  • A. Karthik
Regular Articles


A total of 464 samples comprising of cloacal swabs from necrotic enteritis suspected live birds (191), intestinal scrapings from dead birds with symptoms of necrotic enteritis (91), and apparently healthy birds (182) were collected from selected districts of AP. The samples were subjected to multiplex PCR for simultaneous detection of α, β, and β2 toxin genes and uniplex PCR for the detection of NetB gene. Multiplex PCR screening of samples reveled α toxin gene positives from (cpa) 248/282 (87.94%) necrotic enteritis suspected and 40/182 (21.97%) apparently healthy samples. Among cpa positives 142/248 (57.25%) and 3/40 (7.5%) were positive for β2 toxin gene in necrotic enteritis suspected and apparently healthy birds respectively, indicating the involvement of C. perfringens type A, with minor pore forming toxin gene cpb2 in causing necrotic enteritis in poultry. None of the sample was positive for β toxin gene. The present research indicates C. perfringens type A along with β2 toxin gene was responsible for causing necrotic enteritis in broiler chickens in some parts of Andhra Pradesh in India. Phylogenetic relationship of amplified cpa and cpb2 amino acids sequences from present C. perfringens isolates were studied. The analysis reveals the sequence identity of cpb2 gene of the present isolates and variations at both nucleotide and amino acid level with the published sequences of cpb2 gene of C. perfringens isolates from different animal species of the USA, Iran, Netherlands, and Japan.


Clostridium perfringens Necrotic enteritis Multiplex PCR Phylogenetic analysis Andhra Pradesh 


Funding information

The authors are thankful to Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh, for the financial support for conducting above research.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Veterinary Microbiology, College of Veterinary ScienceSV Veterinary UniversityTirupathiIndia

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