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Chlamydia psittaci is a zoonotic pathogen mainly transmitted by psittacine birds and poultry. The low shedding rate of the pathogen in the apparently healthy birds and human clinical cases may result in false-negative results. In the present study, a droplet digital PCR (ddPCR) assay was developed and compared with optimized quantitative PCR (qPCR) for the detection of C. psittaci from the clinical samples. The ddPCR assay was found to be comparatively more sensitive than the qPCR, wherein the limit of detection (LOD) of ddPCR was upto 2.4 copies of the DNA template, whereas, the qPCR could detect upto 38 copies of the DNA template in the reaction mixture. Overall, the developed ddPCR assay was found to be robust, specific, and could reliably quantify up to 17.8 copies of the DNA template. Finally, the applicability of the developed ddPCR assay was tested by screening the field samples (n = 124), comprising lung tissues from dead poultry and feral birds; pooled faecal samples from the free-living birds, commercial and backyard poultry farms; pharyngeal and cloacal swabs collected from the duck farms. Of these, a total of seven samples were found to be positive by the ddPCR, whereas, three samples could be detected as positive using the qPCR. The developed ddPCR could serve as a reliable screening tool, particularly in those clinical samples wherein the shedding of C. psittaci is substantially very low.

We developed the com1 gene based loop-mediated isothermal amplification (LAMP) assay for the detection of Coxiella burnetii and validated it by screening DNA isolated from serum samples collected from animals and humans. The detection of Coxiella by LAMP assay was comparable with the com1 based-PCR.

A novel Com1 synthetic peptide-based latex agglutination test (LAT) was developed and evaluated against commercial ELISA kit for sero-screening of coxiellosis in cattle. The developed test is economical, has field applicability and can serve as an important rapid tool for sero-screening of coxiellosis in cattle.

Q fever (coxiellosis), caused by Coxiella burnetii, is an emerging or re-emerging zoonotic disease of public health significance and with worldwide distribution. As a causal agent of the one among the 13 global priority zoonoses, having the infectious dose as low as one bacterium, C. burnetii has been regarded as an obligate intracellular bacterial pathogen. The agent has been classified as a Group B bioterrorism agent by the Centre for Disease Control and Prevention (CDC), and the disease is included in the World Organisation for Animal Health (OIE) list of notifiable diseases. It is mainly transmitted through airborne route in humans and animals. Isolation of C. burnetii, using standard routine laboratory culture techniques was impossible until formulation of axenic-based medium. However, it is still to be included among routinely isolated laboratory pathogen, accounting prolonged incubation period (~7 days) and requirement of specific oxygen concentration (2.5% O2). Therefore, indirect diagnostic tools have been mainly used for its diagnosis. So far serology has been mostly used for testing for C. burnetii infection. The detection of C. burnetii DNA by PCR in various clinical samples have also been widely used. The disease has remained largely under-reported, underdiagnosed and as a masked zoonosis; and therefore, needs to be explored through well-planned scientific studies for knowing its true status and likely it impact in humans and animals by employing state-of-the-art diagnostics, identifying its diverse and new host range, as well as risk factors involved in different geo-climatic, behavioural and social settings as well as risk groups. Here, we reviewed the current approaches used for the detection of C. burnetii infection in humans and animals at the population and individual level.