Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Clinical Performance Analysis and Evaluation of Quantitative Real Time PCR for Diagnosis of Scrub Typhus in North East India

Version 1 : Received: 25 May 2020 / Approved: 27 May 2020 / Online: 27 May 2020 (07:51:25 CEST)

How to cite: Paul, D.; Lyngdoh, W.V.; Barman, H.; Lyngdoh, C.J.; Lynrah, K.; Durairaj, E. Clinical Performance Analysis and Evaluation of Quantitative Real Time PCR for Diagnosis of Scrub Typhus in North East India. Preprints 2020, 2020050440. https://doi.org/10.20944/preprints202005.0440.v1 Paul, D.; Lyngdoh, W.V.; Barman, H.; Lyngdoh, C.J.; Lynrah, K.; Durairaj, E. Clinical Performance Analysis and Evaluation of Quantitative Real Time PCR for Diagnosis of Scrub Typhus in North East India. Preprints 2020, 2020050440. https://doi.org/10.20944/preprints202005.0440.v1

Abstract

Scrub typhus is a life-threatening infectious disease and always creating a diagnostic dilemma in terms of rapid turnaround time and accuracy, qRT PCR can become a very good option to achieve the desired result with the molecular level of accuracy and boost up the rapid patient management. This study was performed to evaluate the performance of qRT PCR in comparison to commonly used IgM ELISA and Weil-Felix tests to diagnose scrub typhus, as well as to look for the demographic and clinical profile of the disease in North-East India. It was a hospital-based prospective study conducted in a tertiary care hospital of north-east India, over a period of 1 year, in which all the samples from suspected scrub typhus cases were screened by Weil-Felix test as per institute’s diagnostic protocol after which IgM ELISA for Scrub Typhus was performed. All the IgM positive samples and 20 highly suspected but ELISA negative samples were subjected to qRT PCR, targeting 56 kDa type-specific gene of O. tsutsugamushi. Statistical analysis was done by MS-Excel for Windows v2013® and MedCalc® v17.9 for Windows (MedCalc Software, Acacialaan 22, B-8400 Ostend, Belgium). In this study, we have successfully evaluated the performance of qRT PCR kit for diagnosis of scrub typhus. Out of 54 samples tested, 24 IgM ELISA positive samples and 3 IgM ELISA negative samples have shown the presence of bacterial DNA with quantification of DNA copies. It has also been observed that 21 out of 27 PCR positive samples (77.8%) were detected within the 1st 7 days of illness. All the demographic, as well as clinical data, were also analysed. The performance of the commercial qRT PCR kit used in our study is satisfactory, which provides the extra advantage of quantification of DNA copies and increases diagnostic accuracy within the 1st week of fever.

Keywords

Scrub typhus; qRT PCR; Quantitative PCR; real time PCR; IgM ELISA; North-East India

Subject

Biology and Life Sciences, Immunology and Microbiology

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