|Budget Amount *help
¥2,300,000 (Direct Cost : ¥2,300,000)
Fiscal Year 2003 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 2002 : ¥1,200,000 (Direct Cost : ¥1,200,000)
In this study, we investigated the several problems about antibody detection and nested PCR procedure for more accurate diagnosis of the respiratory tract infection caused by acute Q fever. As for Coxiella antibody detection, five commercial ELISA kits for the diagnosis of acute Q fever.
CIIECKIT-Q-FEVER ELISA, KIT (Bommeli), Q fever IgM / IgG ELISA KIT (Panbio) and Coxiella burnetii ELISA IgM / IgG (Vircell), were examined and compared with IFA titer. For some of the ELISA kits, relatively, good correlation with IFA titer was observed, but it was suggested that cutoff value for of acute infection should be reappraised with the screening of the acute infection in the Japanese population. As for nested PCR, first, four commercial kits and Chelex100 method were compared in its efficacy of DNA extraction. With respiratory tract samples, final detection sensitivity was almost equivalent in any of the DNA extraction methods, while the blood sample showed higher detection sensitivity with the commercial kits clearly than a simple Chelex method. Subsequently, we compared the detection sensitivity and specificity of the several sets of the PCR primers. Nested PCR primers targeted for IS1111a, com1 gene, and 16S r RNA were evaluated, and former 2 primer sets were judged as highly sensitive and specific than 16S primers.