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. 2024 Dec 16;16(12):e75781.
doi: 10.7759/cureus.75781. eCollection 2024 Dec.

Molecular Detection of Tick-Borne Bacterial Pathogens in Patients With Undifferentiated Febrile Illness in India Using a Flow Chip Assay

Chandan K Thakur  1   2 , E V Vinayaraj  2 , Rama Chaudhry  2
Affiliations

Molecular Detection of Tick-Borne Bacterial Pathogens in Patients With Undifferentiated Febrile Illness in India Using a Flow Chip Assay

Chandan K Thakur et al. Cureus. .

Abstract

Background: Tick-borne diseases (TBDs) play a crucial role in human morbidity and mortality, as ticks are highly effective in spreading diseases by transmitting harmful pathogens to humans and animals. The last few decades have seen an increase in the number of recognized tick-borne pathogens and the incidence of TBD worldwide. Several of these diseases are ubiquitous in India. However, in India, there is limited information on the molecular detection of tick-borne pathogens in patients with undifferentiated febrile illness. The study aims to investigate tick-borne pathogens among undifferentiated febrile patients in India using a multiplex polymerase chain reaction (PCR)-based assay.

Methods: A total of 120 blood samples were collected from patients with undifferentiated febrile illnesses of all ages. The samples were tested for a panel of (seven) tick-borne pathogens (Anaplasma, Ehrlichia, Borrelia, Bartonella, Coxiella, Rickettsia, and Francisella) using a multiplex PCR tick-borne bacterial flow chip (TBFC) assay through a semi-automated HybriSpot platform (Vitro Master Diagnóstica, Granada, Spain).

Results: Among the 120 samples tested, one was positive for spotted fever Rickettsia, two were positive for typhus group Rickettsia, one was positive for Borrelia, and two showed coinfections with Borrelia and Anaplasma.

Conclusion: Our knowledge of TBD is steadily advancing with the discovery of novel pathogens and the development of cutting-edge diagnostic tools. Although traditional diagnostic methods like serology and microscopy will remain essential for the diagnosis of TBD, the implementation of advanced molecular diagnostics such as TBFC will enhance our understanding of these diseases by enabling the identification of emerging pathogens and offering more precise and timely diagnoses.

Keywords: febrile illness; hybrispot; rickettsia; tick-borne bacterial flow chip; tick-borne disease; vector; zoonosis.

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Conflict of interest statement

Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. All India Institute of Medical Sciences, New Delhi, Institute Ethics Committee issued approval ref. no.: 214/2018. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: Chandan K. Thakur, E. V. Vinayaraj, Rama Chaudhry declare(s) a grant from Department of Biotechnology, Government of India. The authors gratefully acknowledge the funding provided for this research (Grant No. BT/PR25466/NER/95/1210/2017). Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Upper images show substrate/chromogen reactions producing a dark purple precipitate (hybridization signal) where the PCR amplicon hybridized with the specific probe. Lower corresponding images display signal spots automatically captured and analyzed by the HybriSoft software.
Samples tested positive for (A) Borrelia spp.; (B) Anaplasma phagocytophilum or/and A. bovis or/and A. equi, and Borrelia spp.; (C) Rickettsia typhus group; (D) Rickettsia spotted fever group. PCR: polymerase chain reaction
See this image and copyright information in PMC

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