CLINICAL-EVALUATION OF A NEW PCR ASSAY FOR DETECTION OF COXIELLA-BURNETII IN HUMAN SERUM SAMPLES

A nested PCR method was developed for the detection of Coxiella burnet ii in human serum samples. Two pairs of oligonucleotide primers were d esigned to amplify a 438-bp fragment of the com1 gene encoding a 27-kD a outer membrane protein of C. burnetii. The primers amplified the pre dicted fragments of 21 various strains of C. burnetii but did not reac t with DNA samples from other microorganisms. The 438-bp amplification products could be digested with restriction enzymes SspI and Sail. Th e utility of the nested PCR was evaluated by testing human serum sampl es. The cam1 gene fragment was amplified from 135 (87%) of 155 indirec t immunofluorescence test (IF)-positive serum samples and from 11 (11% ) of 100 IF-negative serum samples. The nested PCR with primers target ed to the com1 gene appeared to be a sensitive, specific, and useful m ethod for the detection of C. burnetii in serum samples.