Comparative analysis of polymerase chain reaction (PCR) assays in detection of Orientia tsutsugamushi from mite vectors
- 1DBT-BiF Centre (Under BTISNet Scheme), Lady Doak College, Madurai, India
Abstract
Orientia tsutsugamushi is the causative agent of scrub typhus infection in humans. This pathogen is transmitted into mammalian hosts including humans through Leptotrombidium mites. Till date, detection of O. tsutsugamushi in human was performed using PCR assay with 56kDa type specific antigen (TSA56), 47kDa outer membrane protein (47kDa-OMP) and groEL chaperonin gene. To understand the efficiency of different genes for the detection of O. tsutsugamushi in Leptotrombidium mites, we performed a comparative PCR assay with various universal primers targeting different genes including TSA56, 47kDa-OMP and groEL gene. We have collected 500 Leptotrombidium mites from Vellore, Tamil Nadu and, the mites were segregated into 50 pools with 10 mites in each pool were used for DNA extraction. The molecular detection of O. tsutsugamushi was performed using PCR assays (including conventional, nested and duplex PCR). The comparative PCR analysis showed nested PCR of groEL gene has high efficiency in detecting the presence of O. tsutsugamushi in chigger samples compared to other genes. Interestingly, groEL target demonstrated high efficiency for detection of O. tsutsugamushi in chigger samples and, when used with nested primers, can facilitate differentiation of O. tsutsugamushi from other rickettsial pathogens. Thus, we conclude groEL gene nested PCR assay can be used routinely to detect O. tsutsugamushi in trombiculid mites.
Related
- Gentamicin Treatment in a Murine Infection Model Suggests Extracellular Orientia tsutsugamushi Contributes to Systemic Dissemination
- Infektion mit Orientia tsutsugamushi und nosokomiale Pneumonie
- Symmetric Inheritance of the Cytoplasmic Bacterium Orientia tsutsugamushi During Mitosis
- First isolation of three Orientia tsutsugamushi strains (Boryong, Kawasaki, and Yonchon) from trombiculid mites in the Republic of Korea
- Prevalence and quantification of viable Campylobacter species in retail chicken: a comparative study using viability digital polymerase chain reaction and conventional culture methods
BibTeX
@misc{govindarajan2026comparative,
title = {{Comparative analysis of polymerase chain reaction (PCR) assays in detection of Orientia tsutsugamushi from mite vectors}},
author = {R. Govindarajan and S. Gowri Sankar and M. Rahi and P. Philip Samuel and A. Alwin Prem Anand},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.28.754975},
url = {https://www.biorxiv.org/content/10.64898/2026.09.28.754975v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Comparative analysis of polymerase chain reaction (PCR) assays in detection of Orientia tsutsugamushi from mite vectors
AU - R. Govindarajan
AU - S. Gowri Sankar
AU - M. Rahi
AU - P. Philip Samuel
AU - A. Alwin Prem Anand
PY - 2026
DA - 2026/09/28
PB - bioRxiv
DO - 10.64898/2026.09.28.754975
UR - https://www.biorxiv.org/content/10.64898/2026.09.28.754975v1
AB - Orientia tsutsugamushi is the causative agent of scrub typhus infection in humans. This pathogen is transmitted into mammalian hosts including humans through Leptotrombidium mites. Till date, detection of O. tsutsugamushi in human was performed using PCR assay with 56kDa type specific antigen (TSA56), 47kDa outer membrane protein (47kDa-OMP) and groEL chaperonin gene. To understand the efficiency of different genes for the detection of O. tsutsugamushi in Leptotrombidium mites, we performed a comparative PCR assay with various universal primers targeting different genes including TSA56, 47kDa-OMP and groEL gene. We have collected 500 Leptotrombidium mites from Vellore, Tamil Nadu and, the mites were segregated into 50 pools with 10 mites in each pool were used for DNA extraction. The molecular detection of O. tsutsugamushi was performed using PCR assays (including conventional, nested and duplex PCR). The comparative PCR analysis showed nested PCR of groEL gene has high efficiency in detecting the presence of O. tsutsugamushi in chigger samples compared to other genes. Interestingly, groEL target demonstrated high efficiency for detection of O. tsutsugamushi in chigger samples and, when used with nested primers, can facilitate differentiation of O. tsutsugamushi from other rickettsial pathogens. Thus, we conclude groEL gene nested PCR assay can be used routinely to detect O. tsutsugamushi in trombiculid mites.
ER -