Tags

Type your tag names separated by a space and hit enter

An ELISA assay using a combination of recombinant proteins from multiple strains of Orientia tsutsugamushi offers an accurate diagnosis for scrub typhus.
BMC Infect Dis. 2017 06 10; 17(1):413.BI

Abstract

BACKGROUND

Scrub typhus (ST) is a disease caused by an obligate intracellular bacterium, Orientia tsutsugamushi, an organism that requires a BSL3 laboratory for propagation. The disease is hallmarked by an eschar at the site of the chigger bite, followed by the development of fever, malaise, myalgia, anorexia, and papulomacular rash. Indirect immunofluorescent assay (IFA) is the gold standard for scrub typhus diagnosis, however, the subjectivity of the assay, the need for a specialized laboratory and instruments has limited the wide use of the test in resource limited areas.

METHODS

A recombinant-protein based enzyme linked immunosorbent assay (ELISA) using the most abundant and immunodominant protein for the detection of Orientia specific antibodies in serum has been developed. The performance of the assay was evaluated using prospectively collected acute sera from 248 randomly selected patients in Thailand. The ELISA assay was evaluated using two different cutoff values.

RESULTS

The receiver operating characteristic (ROC) curve generated cutoff values gave slightly better consistency with diagnosis of ST than those cutoff values established by averaging ELISA optical density of known negatives at 99% confidence interval. Both cutoff values provided similar statistical parameters when compared with the diagnosis of ST, indicating the validity of both calculations to derive cutoff values. These results suggest that both IgG and IgM ELISA performed well to accurately diagnose scrub typhus cases in endemic areas using only acute serum samples.

CONCLUSIONS

We have successfully developed an ELISA assay for the detection of Orientia-specific antibodies in serum that could provide effective screening of acute sera under clinical setup and it is also a useful assay to estimate seroprevalence in various endemic areas.

Authors+Show Affiliations

Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD, USA. chien-chung.c.chao.civ@mail.mil. Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA. chien-chung.c.chao.civ@mail.mil.Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD, USA. Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD, USA. Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA. Department of Medicine, Maharat Nakhon Ratchasima Hospital, Nakhon Ratchasima, Thailand.Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Department of Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Department of Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Department of Medicine, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD, USA. Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

28601091

Citation

Chao, Chien-Chung, et al. "An ELISA Assay Using a Combination of Recombinant Proteins From Multiple Strains of Orientia Tsutsugamushi Offers an Accurate Diagnosis for Scrub Typhus." BMC Infectious Diseases, vol. 17, no. 1, 2017, p. 413.
Chao CC, Zhang Z, Belinskaya T, et al. An ELISA assay using a combination of recombinant proteins from multiple strains of Orientia tsutsugamushi offers an accurate diagnosis for scrub typhus. BMC Infect Dis. 2017;17(1):413.
Chao, C. C., Zhang, Z., Belinskaya, T., Thipmontree, W., Tantibhedyangkul, W., Silpasakorn, S., Wongsawat, E., Suputtamongkol, Y., & Ching, W. M. (2017). An ELISA assay using a combination of recombinant proteins from multiple strains of Orientia tsutsugamushi offers an accurate diagnosis for scrub typhus. BMC Infectious Diseases, 17(1), 413. https://doi.org/10.1186/s12879-017-2512-8
Chao CC, et al. An ELISA Assay Using a Combination of Recombinant Proteins From Multiple Strains of Orientia Tsutsugamushi Offers an Accurate Diagnosis for Scrub Typhus. BMC Infect Dis. 2017 06 10;17(1):413. PubMed PMID: 28601091.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An ELISA assay using a combination of recombinant proteins from multiple strains of Orientia tsutsugamushi offers an accurate diagnosis for scrub typhus. AU - Chao,Chien-Chung, AU - Zhang,Zhiwen, AU - Belinskaya,Tatyana, AU - Thipmontree,Wilawan, AU - Tantibhedyangkul,Wiwit, AU - Silpasakorn,Saowaluk, AU - Wongsawat,Ekkarat, AU - Suputtamongkol,Yupin, AU - Ching,Wei-Mei, Y1 - 2017/06/10/ PY - 2016/11/30/received PY - 2017/06/01/accepted PY - 2017/6/12/entrez PY - 2017/6/12/pubmed PY - 2017/9/26/medline KW - Diagnosis KW - Elisa KW - Recombinant protein antigens KW - Scrub typhus SP - 413 EP - 413 JF - BMC infectious diseases JO - BMC Infect Dis VL - 17 IS - 1 N2 - BACKGROUND: Scrub typhus (ST) is a disease caused by an obligate intracellular bacterium, Orientia tsutsugamushi, an organism that requires a BSL3 laboratory for propagation. The disease is hallmarked by an eschar at the site of the chigger bite, followed by the development of fever, malaise, myalgia, anorexia, and papulomacular rash. Indirect immunofluorescent assay (IFA) is the gold standard for scrub typhus diagnosis, however, the subjectivity of the assay, the need for a specialized laboratory and instruments has limited the wide use of the test in resource limited areas. METHODS: A recombinant-protein based enzyme linked immunosorbent assay (ELISA) using the most abundant and immunodominant protein for the detection of Orientia specific antibodies in serum has been developed. The performance of the assay was evaluated using prospectively collected acute sera from 248 randomly selected patients in Thailand. The ELISA assay was evaluated using two different cutoff values. RESULTS: The receiver operating characteristic (ROC) curve generated cutoff values gave slightly better consistency with diagnosis of ST than those cutoff values established by averaging ELISA optical density of known negatives at 99% confidence interval. Both cutoff values provided similar statistical parameters when compared with the diagnosis of ST, indicating the validity of both calculations to derive cutoff values. These results suggest that both IgG and IgM ELISA performed well to accurately diagnose scrub typhus cases in endemic areas using only acute serum samples. CONCLUSIONS: We have successfully developed an ELISA assay for the detection of Orientia-specific antibodies in serum that could provide effective screening of acute sera under clinical setup and it is also a useful assay to estimate seroprevalence in various endemic areas. SN - 1471-2334 UR - https://www.unboundmedicine.com/medline/citation/28601091/An_ELISA_assay_using_a_combination_of_recombinant_proteins_from_multiple_strains_of_Orientia_tsutsugamushi_offers_an_accurate_diagnosis_for_scrub_typhus_ DB - PRIME DP - Unbound Medicine ER -