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Melioidosis is a life-threatening infectious disease which can lead to high mortality and death within 48 hours if treatment is delayed. Detection of antibodies against Burkholderia pseudomallei may help in early diagnosis of melioidosis. This study was carried out to optimize an in-house ELISA for the detection of Immunoglobulin M (IgM) against B. pseudomallei in suspected acute melioidosis patients in Malaysia. Three different concentrations of crude B. pseudomallei cells antigen (0.200 mg/ml, 0.100 mg/ml and 0.05 mg/ml) and 2 different dilutions of conjugate (1:2000 and 1:4000) were used. The ELISA worked well with 0.05 mg/ml antigen when conjugated with 1:4000 peroxidase-conjugated goat anti-humanIgM. A total of 248 serum samples from 45 culture-confirmed melioidosis patients, 117 non-melioidosis patients (leptospirosis (n = 18), tuberculosis (n = 8), rheumathoid arthritis (n = 37), dengue (n = 12), hepatitis B (n = 23) and chikungunya (n = 19), and 86 healthy blood donors were used to construct receiver operating characteristics (ROC) curves. Based on the ROC analysis, this in-house IgM ELISA showed the highest sensitivity and specificity up to 82.2% and 94.87% respectively at the cut-off 0.605 serum absorbance. At this cut-off value, only 6 false positives were detected among 117 non-melioidosis patients, and the remaining 111 were detected as true negative. In conclusion, this in-house IgM ELISA can be applied for the laboratory diagnosis of acute melioidosis in Malaysian hospitals.

The Optimization of Igm In-House ELISA for the Laboratory Diagnosis of Melioidosis in Malaysia