Genotyping of Indian antigenic, vaccine, and field Brucella spp. using multilocus sequence typing

  • Rajeswari Shome ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
  • Natesan Krithiga ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
  • Padmashree B Shankaranarayana ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
  • Sankarasubramanian Jegadesan School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India
  • Vishnu Udayakumar S School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India
  • Bibek Ranjan Shome ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
  • Girin Kumar Saikia College of Veterinary Sciences, Assam Agricultural University, Khanapara, Guwahati, Assam, India
  • Narendra Kumar Sharma Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India
  • Harshad Chauhan College of Veterinary Science and Animal Husbandry, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India
  • Bharat Singh Chandel College of Veterinary Science and Animal Husbandry, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India
  • Rajendhran Jeyaprakash School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India
  • Habibur Rahman ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India

Abstract

Introduction: Brucellosis  is one of the most important zoonotic diseases that affects  multiple livestock species and causes great economic losses. The highly conserved genomes of Brucella, with > 90% homology among species, makes it important to study the genetic diversity circulating in the country.

Methodology: A total of 26 Brucella spp. (4 reference strains and 22 field isolates) and 1 B. melitensis draft genome sequence from India (B. melitensis Bm IND1) were included for sequence typing. The field isolates were identified by biochemical tests and confirmed by both conventional and quantitative polymerase chain reaction (qPCR) targeting bcsp 31Brucella genus-specific marker. Brucella speciation and biotyping was done by Bruce ladder, probe qPCR, and AMOS PCRs, respectively, and genotyping was done by multilocus sequence typing (MLST).

Results: The MLST typing of 27 Brucella spp. revealed five distinct sequence types (STs); the B. abortus S99 reference strain and 21 B. abortus field isolates belonged to ST1. On the other hand, the vaccine strain B. abortus S19 was genotyped as ST5. Similarly, B. melitensis 16M reference strain and one B. melitensis field isolate were grouped into ST7. Another B. melitensis field isolate belonged to ST8 (draft genome sequence from India), and only B. suis 1330 reference strain was found to be ST14.

Conclusion: The sequences revealed genetic similarity of the Indian strains to the global reference and field strains. The study highlights the usefulness of MLST for typing of field isolates and validation of reference strains used for diagnosis and vaccination against brucellosis.

Author Biographies

Rajeswari Shome, ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
Bacteriology Section
Natesan Krithiga, ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
Bacteriology Section
Padmashree B Shankaranarayana, ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
Bacteriology Section
Sankarasubramanian Jegadesan, School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Dept of Genetics
Vishnu Udayakumar S, School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Dept of genetics
Bibek Ranjan Shome, ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
Bacteriology Section
Girin Kumar Saikia, College of Veterinary Sciences, Assam Agricultural University, Khanapara, Guwahati, Assam, India

Dept. of Microbiology, College of Veterinary Sciences, Assam Agricultural University
Narendra Kumar Sharma, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India
Senior Bacteriologist cum Head
Deptt of Veterinay Microbiology
Guru Angad Dev Veterinary and Animal Sciences University
Harshad Chauhan, College of Veterinary Science and Animal Husbandry, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India

Dept. of Animal Biotechnology, College of Veterinary Science and Animal Husbandry
Bharat Singh Chandel, College of Veterinary Science and Animal Husbandry, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India

Dept. of Animal Biotechnology, College of Veterinary Science and Animal Husbandry,
Sardarkrushinagar Dantiwada Agricultural University
Rajendhran Jeyaprakash, School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, India

Dept. of Genetics, School of Biological Sciences, Madurai Kamaraj University
Habibur Rahman, ICAR - National Institute of Veterinary Epidemiology and Disease Informatics (ICAR - NIVEDI) (Formerly PD_ADMAS),Yelahanka, Bengaluru, Karnataka, India
Bacteriology section
Published
2016-03-31
How to Cite
Shome R, Krithiga N, Shankaranarayana P, Jegadesan S, Udayakumar S V, Shome B, Saikia G, Sharma N, Chauhan H, Chandel B, Jeyaprakash R, Rahman H (2016) Genotyping of Indian antigenic, vaccine, and field Brucella spp. using multilocus sequence typing. The Journal Of Infection In Developing Countries 10 (03): 237-244. https://doi.org/10.3855/jidc.6617
Section
Original Articles

Keywords

Brucella; AMOS PCR; Bruce PCR; Indian isolates; MLST; qPCR