Vet World Vol.12 November-2019 Article-11
Research Article
Veterinary World, 12(11): 1769-1774
https://doi.org/10.14202/vetworld.2019.1769-1774
Application of immunofluorescence assay and nested polymerase chain reaction for query fever diagnosis in animal handlers of Puducherry, South India, and phylogenetic analysis based on IS1111 repetitive gene element
2. Department of Veterinary Microbiology, Rajiv Gandhi Institute of Veterinary Education and Research, Puducherry, India.
3. Central Interdisciplinary Research Facility, Mahatma Gandhi Medical College and Research Institute, Sri Balaji Vidyapeeth (Deemed to be University), Puducherry, India.
Background and Aim: Diagnosis of query fever (QF) is mostly done on the basis of serological/molecular tests, due to the stringent requirement of biosafety level-3 containment facilities for isolating Coxiella burnetii in culture. QF is an important zoonosis and is considered to be an occupational hazard to livestock handlers. This report describes our study on the serological as well as molecular evidence of QF in animal handlers from Puducherry and surrounding Tamil Nadu, from where, to the best of our knowledge, no such reports are available so far.
Materials and Methods: Seventy-five animal handlers were recruited, comprising veterinarians, slaughterhouse workers, butchers, and animal attendants of various government veterinary clinics from Puducherry and surrounding areas of Tamil Nadu state. QF serology was performed to identify Phase I and Phase II immunoglobulin G antibodies to C. burnetii. Nested polymerase chain reaction (N-PCR) was carried out to detect C. burnetii DNA in buffy coat samples by targeting IS1111 gene element. N-PCR-positive samples were sequenced and phylogenetic analysis was performed using MEGA software version 10.0.
Results: A total of 21 animal handlers (28.1%) were positive for either serology or PCR. PCR alone was positive in 10 (13.4%), only serology was positive in 8 (10.7%), and both serology and PCR were positive in three samples (4.0%). GenBank accession numbers were obtained for 13 N-PCR-positive samples (MG548608-MG548620). Six of our study sequences showed close similarity with the reference isolates from Bengaluru, Colombia, Brazil, France, and Iran.
Conclusion: A significant percentage of QF positivity in animal handlers of this part of South India, Puducherry, warrants a prospective study with follow-up of a large number of this occupational group. Keywords: Coxiella burnetii, immunofluorescence assay immunoglobulin G, Q fever in Puducherry, query fever nested polymerase chain reaction, zoonosis.
Keywords: Coxiella burnetii, immunofluorescence assay immunoglobulin G, Q fever in Puducherry, query fever nested polymerase chain reaction, zoonosis.
How to cite this article: Pradeep J, Stephen S, Sangeetha B, Antony PX, Amsaveni S, Pooja P (2019) Application of immunofluorescence assay and nested polymerase chain reaction for query fever diagnosis in animal handlers of Puducherry, South India, and phylogenetic analysis based on IS1111 repetitive gene element, Veterinary World, 12(11): 1769-1774.
Received: 23-07-2019 Accepted: 09-10-2019 Published online: 13-11-2019
Corresponding author: Selvaraj Stephen E-mail: stephens4950@gmail.com
DOI: 10.14202/vetworld.2019.1769-1774
Copyright: Pradeep, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.