title: Evaluation of PCR-based assay for diagnosis of spotted fever group rickettsiosis in human serum samples.
abstract: A nested PCR assay was developed for the detection of spotted fever group (SFG) rickettsiae in serum samples. The assay was based on specific primers derived from the rickettsial outer membrane protein B gene (rompB) of Rickettsia conorii. An SFG rickettsia-specific signal is obtained from R. akari, R. japonica, R. sibirica, and R. conorii. Other bacterial species tested did not generate any signal, attesting to the specificity of the assay. As few as seven copies of the rompB gene of R. conorii could be detected in 200 microl of serum sample. The assay was evaluated with a panel of sera obtained from patients with acute-phase febrile disease tested by immunofluorescent antibody assay (IFA). The SFG rickettsia-specific DNA fragment was detected in 71 out of 100 sera, which were proven to have immunoglobulin M antibodies against SFG rickettsial antigen by IFA. The results were further confirmed by restriction fragment length polymorphism and sequencing analysis of the DNA fragments. The results indicated that this PCR assay is suitable for the diagnosis of spotted fever group rickettsiosis in Korea.
authors.affiliation: Department of Microbiology, College of Medicine, Konkuk University, Choongu-shi, Choongbuk 380-701, Republic of Korea.
authors.forename: Yeon-Joo
authors.initials: YJ
authors.lastname: Choi
authors.forename: Seung-Hyun
authors.initials: SH
authors.lastname: Lee
authors.forename: Kyung-Hee
authors.initials: KH
authors.lastname: Park
authors.forename: Young-Sang
authors.initials: YS
authors.lastname: Koh
authors.forename: Keun-Hwa
authors.initials: KH
authors.lastname: Lee
authors.forename: Hyung-Suk
authors.initials: HS
authors.lastname: Baik
authors.forename: Myung-Sik
authors.initials: MS
authors.lastname: Choi
authors.forename: Ik-Sang
authors.initials: IS
authors.lastname: Kim
authors.forename: Won-Jong
authors.initials: WJ
authors.lastname: Jang
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000097:blood* / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000097:blood* / Q000235:genetics; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D006801:Humans; D007723:Korea / Q000453:epidemiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012281:Rickettsia / Q000145:classification / Q000235:genetics*; D012282:Rickettsia Infections / Q000175:diagnosis* / Q000378:metabolism
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C073150:rOmp B protein, Rickettsia