Coxiellosis in domestic livestock of Puducherry and Tamil Nadu: Detection of Coxiella burnetii DNA by polymerase chain reaction in slaughtered ruminants

Abstract

Background and aim: In the course of our Indian Council of Medical Research project on coxiellosis in Puducherry and Tamil Nadu, 5.64% goat, 1.85% sheep, 1.06% buffaloes, and 0.97% cattle were positive for Coxiella burnetii antibodies by enzyme linked immunosorbent assay kit (IDEXX, Liebefeld, Switzerland). In this preliminary study, we have proceeded to look for C. burnetii DNA in those antibody positive specimens employing an imported commercial C. burnetii polymerase chain reaction (PCR) kit.

Materials and methods: Blood samples were collected during slaughtering. All 15 blood samples of antibody positive ruminants and three antibody negative samples were subjected to conventional Trans-PCR assay with a commercial PCR kit (Genekam Biotechnology AG, Duisburg, Germany). An in-house Trans-PCR was included in the study for comparison.

Results: A total of 15 antibody positive and three antibody-negative serum samples belonging to 11 goat, 4 sheep, 1 cattle, and 2 buffaloes were tested in duplicate for the presence of C. burnetii DNA by the commercial agar gel PCR kit and an in-house Trans-PCR. Only one buffalo serum sample was positive for C. burnetii with a band at 243 bp in in-house Trans-PCR.

Discussion: Seropositivity for C. burnetii need not necessarily translate into infectivity status of the animal. Conversely, seronegative ruminants can shed C. burnetii. Rapid disintegration of C. burnetii DNA during the storage period is an important impediment in QF-PCR research. This is the first time the performance of this commercial PCR kit is being validated in India.

Conclusion: Commercial PCR kit, Genekam did not identify any positive sample, probably because it targeted a larger amplicon of 687 bp.

Keywords: Coxiella burnetii DNA; Trans-polymerase chain reaction; coxiellosis.

Figure-1

Coxiella burnetii Genekam conventional polymerase chain reaction. m – DNA ladder 100 bp; 2, 4, 7, 13, 14, 17, 19, 20, 33, 36 – Negative; ntc – negative control; pc – positive control (687 bp).

Figure-2

Coxiella burnetii in-house Trans-polymerase chain reaction (Trans 3 and 4 primers). m – DNA ladder 100 bp; 2, 4, 7, 14, 17, 19, 20, 33, 36 – Negative; 13 – Positive for C. brunetti DNA; ntc – negative control; pc – positive control (243 bp).