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Clinical Utility of Combined Whole-cell Antigen and Recombinant Hemolysis Co-regulated Protein 1-Enzyme-linked Immunosorbent Assays Reveals Underdiagnosed Cases of Melioidosis in Vietnam

Quyen T. L. TranVNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam;

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Ha V. NguyenVNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam;

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Huyen T. PhamGeneral Hospital of Ha Tinh Province, Ha Tinh, Vietnam;

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Tuan V. MaiHue Central Hospital, Hue, Vietnam

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Quyen H. M. NguyenVNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam;

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Dzung V. LeGeneral Hospital of Ha Tinh Province, Ha Tinh, Vietnam;

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Linh N. H. BuiVNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam;

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Lan T. H. HoangHue Central Hospital, Hue, Vietnam

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Trung Q. HoangGeneral Hospital of Ha Tinh Province, Ha Tinh, Vietnam;

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Trung T. TrinhVNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam;

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ABSTRACT.

Melioidosis is a fatal infectious disease in the tropics and subtropics. Currently, bacterial culture is the gold standard for diagnosis of the disease, but its sensitivity is relatively low. In this study, we evaluated four ELISAs using sera collected from culture-confirmed cases of melioidosis (n = 63), cases with other bacterial infections (n = 62), and healthy donors (n = 60). Antigens used for ELISAs were the whole-cell (WC) antigens and recombinant proteins of hemolysis co-regulated protein 1 (Hcp1), chaperonin GroEL1, and alkyl hydroperoxide reductase subunit C (AhpC). Using the cutoff values for optical density at 490 nm defined at a specificity of > 95%, the sensitivity of the WC, Hcp1, GroEL1, and AhpC ELISAs was 93.7%, 87.3%, 61.9%, and 57.1%, respectively. The combined WC/Hcp1 ELISA showed the greatest sensitivity and specificity of 98.4% and 95.1%, respectively. Of 511 and 500 sera collected from clinically suspected febrile patients admitted to the General Hospital of Ha Tinh Province and the Hue Central Hospital, respectively, combined WC/Hcp1 ELISAs showed 52 (10.2%) and 41 (8.2%) patients positive for melioidosis, respectively. The assay detected 14 of 14 (100%) and 21 of 23 (91.3%) culture-confirmed cases of melioidosis at Ha Tinh and Hue, respectively. A follow-up study of 38 patients positive for melioidosis by combined WC/Hcp1 ELISAs but negative for Burkholderia pseudomallei by culture method or not assigned to examine for bacterial culture resulted in 2 (5.3%) culture-reconfirmed patients with melioidosis, 9 (23.7%) deaths, 17 (44.7%) unhealthy patients, and 10 (26.3%) healthy persons. Combined WC/Hcp1 ELISA was a reliable serological method to detect underdiagnosed cases of melioidosis. Further investigations are needed to estimate the true sensitivity and specificity of the assay and the true number of cases of melioidosis.

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Author Notes

Address correspondence to Trung T. Trinh, Laboratory for Microbial Pathogens, Institute of Microbiology and Biotechnology, Vietnam National University, 144 Xuan Thuy, Cau Giay, Hanoi, Vietnam. E-mail: tttrung@vnu.edu.vn

Financial support: This study was funded by the Ministry of Science and Technology in Vietnam (grant no. NDT.82.GB/20).

Authors’ addresses: Quyen T. L. Tran, Ha V. Nguyen, Quyen H. M. Nguyen, Linh N. H. Bui, and Trung T. Trinh, VNU-Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam, E-mails: quyenttl@vnu.edu.vn, ha.nguyenviet310@gmail.com, quyennhm@vnu.edu.vn, linhbui299@vnu.edu.vn, and tttrung@vnu.edu.vn. Huyen T. Pham, Dzung V. Le, and Trung Q. Hoang, General Hospital of Ha Tinh Province, Ha Tinh, Vietnam, E-mails: huyenpham.bvdkht@gmail.com, dzungtmht@gmail.com, and hoangquangtrung.bs@gmail.com. Tuan V. Mai and Lan T. H. Hoang, Hue Central Hospital, Hue, Vietnam, E-mails: maituanbvh@yahoo.com.vn and hglanhuong.hch@gmail.com.

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