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Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus.
J Clin Microbiol. 2017 05; 55(5):1377-1387.JC

Abstract

Scrub typhus, caused by Orientia tsutsugamushi, is a common cause of acute undifferentiated febrile illness in the Asia-Pacific region. However, its nonspecific clinical manifestation often prevents early diagnosis. We propose the use of PCR and serologic tests as diagnostic tools. Here, we developed a multiplex real-time PCR assay using hydrolysis (TaqMan) probes targeting O. tsutsugamushi 47-kDa, groEL, and human interferon beta (IFN-β gene) genes to improve early diagnosis of scrub typhus. The amplification efficiency was higher than 94%, and the lower detection limit was 10 copies per reaction. We used a human gene as an internal DNA quality and quantity control. To determine the sensitivity of this PCR assay, we selected patients with confirmed scrub typhus who exhibited a clear 4-fold increase in the level of IgG and/or IgM. The PCR assay result was positive in 45 of 52 patients, indicating a sensitivity of 86.5% (95% confidence interval [CI]: 74.2 to 94.4). The PCR assessment was negative for all 136 non-scrub typhus patients, indicating a specificity of 100% (95% CI: 97.3 to 100). In addition, this test helped diagnose patients with inconclusive immunofluorescence assay (IFA) results and using single blood samples. In conclusion, the real-time PCR assay proposed here is sensitive and specific in diagnosing scrub typhus. Combining PCR and serologic tests will improve the diagnosis of scrub typhus among patients presenting with acute febrile illness.

Authors+Show Affiliations

Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand wiwit.tan@mahidol.ac.th yupin.sup@mahidol.ac.th.Division of Infectious Diseases and Tropical Medicine, Department of Internal Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Division of Infectious Diseases and Tropical Medicine, Department of Internal Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Division of Infectious Diseases and Tropical Medicine, Department of Internal Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Maharat Nakhon Ratchasima Hospital, Nakhon Ratchasima Province, Thailand.Division of Infectious Diseases and Tropical Medicine, Department of Internal Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand wiwit.tan@mahidol.ac.th yupin.sup@mahidol.ac.th.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28202789

Citation

Tantibhedhyangkul, Wiwit, et al. "Use of Multiplex Real-Time PCR to Diagnose Scrub Typhus." Journal of Clinical Microbiology, vol. 55, no. 5, 2017, pp. 1377-1387.
Tantibhedhyangkul W, Wongsawat E, Silpasakorn S, et al. Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus. J Clin Microbiol. 2017;55(5):1377-1387.
Tantibhedhyangkul, W., Wongsawat, E., Silpasakorn, S., Waywa, D., Saenyasiri, N., Suesuay, J., Thipmontree, W., & Suputtamongkol, Y. (2017). Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus. Journal of Clinical Microbiology, 55(5), 1377-1387. https://doi.org/10.1128/JCM.02181-16
Tantibhedhyangkul W, et al. Use of Multiplex Real-Time PCR to Diagnose Scrub Typhus. J Clin Microbiol. 2017;55(5):1377-1387. PubMed PMID: 28202789.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus. AU - Tantibhedhyangkul,Wiwit, AU - Wongsawat,Ekkarat, AU - Silpasakorn,Saowaluk, AU - Waywa,Duangdao, AU - Saenyasiri,Nuttawut, AU - Suesuay,Jintapa, AU - Thipmontree,Wilawan, AU - Suputtamongkol,Yupin, Y1 - 2017/02/15/ PY - 2016/10/27/received PY - 2017/02/06/accepted PY - 2017/2/17/pubmed PY - 2017/11/1/medline PY - 2017/2/17/entrez KW - Orientia tsutsugamushi KW - real-time PCR KW - scrub typhus SP - 1377 EP - 1387 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 55 IS - 5 N2 - Scrub typhus, caused by Orientia tsutsugamushi, is a common cause of acute undifferentiated febrile illness in the Asia-Pacific region. However, its nonspecific clinical manifestation often prevents early diagnosis. We propose the use of PCR and serologic tests as diagnostic tools. Here, we developed a multiplex real-time PCR assay using hydrolysis (TaqMan) probes targeting O. tsutsugamushi 47-kDa, groEL, and human interferon beta (IFN-β gene) genes to improve early diagnosis of scrub typhus. The amplification efficiency was higher than 94%, and the lower detection limit was 10 copies per reaction. We used a human gene as an internal DNA quality and quantity control. To determine the sensitivity of this PCR assay, we selected patients with confirmed scrub typhus who exhibited a clear 4-fold increase in the level of IgG and/or IgM. The PCR assay result was positive in 45 of 52 patients, indicating a sensitivity of 86.5% (95% confidence interval [CI]: 74.2 to 94.4). The PCR assessment was negative for all 136 non-scrub typhus patients, indicating a specificity of 100% (95% CI: 97.3 to 100). In addition, this test helped diagnose patients with inconclusive immunofluorescence assay (IFA) results and using single blood samples. In conclusion, the real-time PCR assay proposed here is sensitive and specific in diagnosing scrub typhus. Combining PCR and serologic tests will improve the diagnosis of scrub typhus among patients presenting with acute febrile illness. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/28202789/Use_of_Multiplex_Real_Time_PCR_To_Diagnose_Scrub_Typhus_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=28202789 DB - PRIME DP - Unbound Medicine ER -