Notably, selleck chemical GroEL

had the highest sensitivity and modest specificity for Momelotinib mouse recognizing of Q fever, which may be the most important antigen used for the diagnosis of Q fever. The antigen combination, GroEL, YbgF and Com1, may give a more authentic specificity. Refinement of antigen combination and the production of fusion molecules comprised of the major seroreactive antigens described herein may lead to improved sensitivity and specificity for the development of a rapid, accurate, and convenient seorodiagnostic test of Q fever. Conclusions In summary, the combination of 2D-PAGE, immunoblot and MALDI-TOF-MS permitted the identification of 20 seroreactive proteins of C. burnetii. A protein microarray fabricated Selleckchem NVP-BGJ398 with recombinant proteins was probed with Q fever

patient sera. Seven proteins (GroEL, YbgF, RplL, Mip, Com1, OmpH, and Dnak) were recognized as major seroreactive antigens. The major seroreactive proteins fabricated in a small array were analyzed with the sera of patients with Q fever, rickettsial spotted fever, Legionella pneumonia or streptococcal pneumonia and they gave a moderate specificity for recognizing of Q fever patient sera, suggesting these proteins are potential serodiagnostic markers for Q fever. Methods Culture and purification of C. burnetii C. burnetii Xinqiao strain (phase I) was propagated in embryonated eggs and purified by renografin density centrifugation as previously described [25]. The purified organisms were suspended in phosphate-buffered saline buffer (PBS) (8.1 mM Na2HPO4, 1.9 mM NaH2PO4, 154 mM NaCl, PH7.4) and stored at −70°C. Mouse and human sera Thirty two BALB/c mice (male, 6 weeks Thymidylate synthase old) (Laboratory Animal Center of Beijing, China) were injected intraperitoneally with C. burnetii Xinqiao strain (1 × 108 cells/mouse) in a biosafety level 3 laboratory. Eight of the mice were randomly sacrificed on days 7, 14, 21, and 28 pi. Ten mg of tissue from the liver, spleen and lungs of each sacrificed mouse was used to extract DNA with a tissue DNA extraction kit (Qiagen, GmbH, Germany), respectively. Each DNA sample was eluted from the DNA extraction column with 200 μl elution buffer according

to the manufacturer’s instruction. A 2 μl of the DNA sample was tested by a real-time quantitative polymerase chain reaction (qPCR) specific for C. burnetii [26]. The results of qPCR were expressed as mean ± SD and compared by the repeated measurement data analysis of variance using SAS 9.1 software (SAS Institute Inc., Cary, NC). All animal protocols were pre-approved by the Animal Protection Committee of Laboratory Animal Center of Beijing and all experiments complied with the current laws of China. Fifty six serum samples from Q fever patients were obtained from the Australian Rickettsial Reference Laboratory (Geelong, VIC, Australia) and classified into 3 types, acute early, acute late and convalescent according to the results of the IFA results and clinical details of the patients.