Use of Recombinant Escherichia coli Strains in Immunofluorescence Assays for Melioidosis Diagnosis

Burkholderia pseudomallei is a Gram-negative bacterium and the causative agent of melioidosis in humans and animals in the tropics. The clinical manifestations of melioidosis are diverse, ranging from localized infections to whole-body sepsis. The effective serological method is crucial for the poin...

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Bibliographic Details
Published in:Pathogens (Basel) 2021-05, Vol.10 (5), p.559
Main Authors: Lantong, Kanoknart, Songsri, Jirarat, Wisessombat, Sueptrakool, Mala, Wanida, Prommachote, Warinda, Senghoi, Wilaiwan, Kotepui, Manas, Kaewrakmuk, Jedsada, Jiranantasak, Treenate, Tuanyok, Apichai, Klangbud, Wiyada Kwanhian
Format: Article
Language:English
Subjects:
Accuracy
Antibodies
Antigens
Bacteria
Burkholderia pseudomallei
Cell culture
Cross-reactivity
Diagnosis
E coli
Efficiency
Escherichia coli
Hemagglutination
IgG antibody
Immunofluorescence
immunofluorescence assay
Immunoglobulin G
Indirect hemagglutination
Laboratories
Melioidosis
Motility
Proteins
Sepsis
Serology
Tropical environments
TssM
whole-cell-based assay
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Summary:Burkholderia pseudomallei is a Gram-negative bacterium and the causative agent of melioidosis in humans and animals in the tropics. The clinical manifestations of melioidosis are diverse, ranging from localized infections to whole-body sepsis. The effective serological method is crucial for the point-of-care diagnosis of melioidosis. The aim of this study was to develop indirect immunofluorescence assay (IFA)-based methods for detecting immunoglobulin G (IgG) antibodies in melioidosis patients. These methods use whole-cell antigens made from recombinant E. coli strains that express major B. pseudomallei antigens, including TssM, OmpH, AhpC, BimA, and Hcp1. A total of 271 serum samples from culture-confirmed melioidosis patients (n = 81), patients with other known infections (n = 70), and healthy donors (n = 120) were tested. Our study showed that the recombinant TssM strain had the highest performance, with 92.6% sensitivity, 100% specificity, 100% positive predictive value, 96.9% negative predictive value, 97.8% efficiency, 97.0% accuracy, and no cross-reactivity. The method agreement analysis based on k efficiency calculations showed that all five IFA methods perfectly agreed with the standard culturing method, while the traditional indirect hemagglutination (IHA) method moderately agreed with the culture. In summary, our investigations showed that the TssM-IFA method could be used for melioidosis diagnosis.
ISSN:2076-0817
2076-0817
DOI:10.3390/pathogens10050559