Serological Diagnosis of Human Melioidosis with Indirect Hemagglutination and Complement Fixation Tests

Abstract

An indirect hemagglutination (IHA) test and a complement fixation (CF) test were evaluated from test results on sera from 212 human melioidosis patients of which 119 were culturally proved cases. Significant antibody titers (IHA titers of 1:40 or greater and CF titers of 1:4 or greater) were demonstrated with either test in all except five patients. IHA and CF titers ranged as high as 1:20,480 and 1:1,024, respectively. Antibodies were usually demonstrated by both tests 1 week after onset of disease. Transient seronegative reactions during the course of disease were seen in sera of approximately 19% of the patients with either IHA and CF but rarely with both tests. High titers in either test were obtained by the third week of disease and reached maximum levels in 4 to 5 months. Titers usually were detectable for 9 or more months. Antibodies were detected by IHA and CF tests in 80 to 100% of the sera obtained at various time intervals from 9 months to 2 or more years after disease onset. Antibody persistence occurred in patients who had a short disease course, as well as in patients with prolonged, complicated infections. The IHA test had excellent specificity when evaluated with normal human sera and diverse antimicrobial sera from hyperimmunized rabbits and human patients. The CF antigen appeared to contain common antigens with some but not all types of Pseudomonas aeruginosa. The specificity of the CF antigen could be enhanced without appreciable effect on its sensitivity by use of a titer of 1:8 in lieu of 1:4 as a criterion for a significant reaction. Either test could be used advantageously for the laboratory diagnosis of melioidosis.

Human Immune Responses to Melioidosis and Cross-Reactivity to Low-Virulence Burkholderia Species, Thailand1

“…We prepared antigens in accordance with published methods, unless otherwise stated (17,18). For IHA, we used crude culture filtrate antigens from pooled isolates with the following specifications: B. pseudomallei Thai clinical isolates 199a and 207a (19), B. thailandensis Thai environmental isolates E264 (ATCC700388) and STBCC006 (20), and BTCV Thai environmental E555 and USA clinical isolate CDC3015869.…”

Prevalence of Antibodies to Pseudomonas pseudomallei Exotoxin and Whole Cell Antigens in Military Personnel in Sabah and Sarawak, Malaysia

“…Furthermore, Nigg and Johnson (11) reported that P. aeruginosa antigen was shown to be capable of cross-reacting with antibodies to P. pseudomallei surface antigens when complement-fixation serologic technique was used. Similarly, cross-reactivity between antigens of P. pseudomallei and P. stutzeri was also evident when complement-fixation procedure was employed by Alexander et al (1). Exotoxins of P. pseudomallei are proteins of relatively lower molecular weight (36,000 daltons) compared to that of P. aeruginosa exotoxin (M.W.…”

Polysaccharide microarray technology for the detection of Burkholderia pseudomallei and Burkholderia mallei antibodies

“…In the present study, we are able to demonstrate the usefulness of polysaccharide microarray technology for the detection of serum antibodies against B. mallei and B. pseudomallei. An indirect hemagglutination and a complement fixation (CF) test had been the routine procedure for the serologic diagnosis of human melioidosis caused by B. pseudomallei infection (Alexander et al, 1970;Wuthiekanum et al, 2006). B. mallei infections (glanders) in horses had been serologically diagnosed employing CF test (Marek and Manninger, 1945).…”