A Rapid Immunochromatography Test Based on Hcp1 Is a Potential Point-of-Care Test for Serological Diagnosis of Melioidosis

Phokrai, Phornpun; Karoonboonyanan, W; Thanapattarapairoj, Nida; Promkong, Chidchanok; Dulsuk, Adul; Koosakulnirand, Sirikamon; Canovali, Sasha; Indrawattana, Nitaya; Jutrakul, Yaowaruk; Wuthiekanun, Vanaporn; Limmathurotsakul, Direk; Brett, Paul J.; Burtnick, Mary N.; Lertmemongkolchai, Ganjana; Chantratita, Narisara

doi:10.1128/jcm.00346-18

Cited by 37 publications

(53 citation statements)

References 22 publications

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“…To combine the advantages of a multiplex approach with a rapid test POC format we decided to develop a 4-plex dipstick (“DS”) assay using the three best performing antigens BPSL2096, BPSL2697, BPSS0477 of our previously published protein microarray [ 32 ] and the hemolysin-coregulated protein (Hcp1) BPSS1498. The latter was included as recent studies by Chantratita and colleagues reported a strong serological discrimination power for this antigen [ 31 , 37 ]. The selected antigens included both B .…”

Section: Resultsmentioning

confidence: 99%

“…We decided to detect human IgG against B . pseudomallei antigens as a readout for our assay, as it has been shown to be a valuable diagnostic marker in serological melioidosis assays [ 31 , 39 – 41 ]. Although IgM detection has also been applied previously, no clear benefit over IgG based assays has been shown.…”

Section: Resultsmentioning

confidence: 99%

“…The four serodiagnostic protein antigens BPSL2096, BPSL2697, BPSS0477 and BPSS1498 (see Table 1 ) were selected based on their sensitivity and specificity published previously by others and us [ 31 , 32 ].…”

Section: Methodsmentioning

confidence: 99%

“…We also tested another 95 samples from Thailand on our dipsticks. These sera were previously classified as false-negative (n = 55) or false-positive (n = 40) on an Hcp1 based lateral flow assay [ 31 ]. Twenty-eight of the false-positive sera were drawn from healthy individuals and 12 from patients suffering from other kinds of infections.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, a new generation of serological assays attracted great attention [ 31 , 32 ]. A rapid immunochromatography test based on the hemolysin-coregulated protein (Hcp1) with high sensitivity and specificity shows great promise as a point of care (POC) assay, being well standardized and inexpensive [ 31 ].…”

Section: Introductionmentioning

confidence: 99%

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Melioidosis DS rapid test: A standardized serological dipstick assay with increased sensitivity and reliability due to multiplex detection

et al. 2020

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Background Melioidosis, caused by Burkholderia pseudomallei , is a severe infectious disease with high mortality rates, but is under-recognized worldwide. In endemic areas, there is a great need for simple, low-cost and rapid diagnostic tools. In a previous study we showed, that a protein multiplex array with 20 B . pseudomallei -specific antigens detects antibodies in melioidosis patients with high sensitivity and specificity. In a subsequent study the high potential of anti- B . pseudomallei antibody detection was confirmed using a rapid Hcp1 single protein-based assay. Our protein array also showed that the antibody profile varies between patients, possibly due to a combination of host factors but also antigen variations in the infecting B . pseudomallei strains. The aim of this study was to develop a rapid test, combining Hcp1 and the best performing antigens BPSL2096, BPSL2697 and BPSS0477 from our previous study, to take advantage of simultaneous antibody detection. Methods and principal findings The 4-plex dipstick was validated with sera from 75 patients on admission plus control groups, achieving 92% sensitivity and 97–100% specificity. We then re-evaluated melioidosis sera with the 4-plex assay that were previously misclassified by the monoplex Hcp1 rapid test. 12 out of 55 (21.8%) false-negative samples were positive in our new dipstick assay. Among those, 4 sera (7.3%) were Hcp1 positive, whereas 8 (14.5%) sera remained Hcp1 negative but gave a positive reaction with our additional antigens. Conclusions Our dipstick rapid test represents an inexpensive, standardized and simple diagnostic tool with an improved serodiagnostic performance due to multiplex detection. Each additional band on the test strip makes a false-positive result more unlikely, contributing to its reliability. Future prospective studies will seek to validate the gain in sensitivity and specificity of our multiplex rapid test approach in different melioidosis patient cohorts.

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Section: Resultsmentioning

confidence: 99%