RESUMEN
Objective To study the different proteins of human and environment Vibrio cholerae (V. cholerae) by two dimensional electrophoresis (2-DE) and TOF-TOF-MS. Methods Total cellular pro-tein was extracted by lysate from V. cholerae, and proteins were separated by 2-DE under immobilized pH gradients(IPG), then electrophoregrams were dealed with coomassie brilliant blue, and analyzed by Im-ageMaster 2D Elite 5.0, finally the different protein spots were identified by TOF-TOF-MS. Results High repetitive 2-DE maps were obtained. 2-DE and image analysis revealed 1032±22 protein spots, and PI value was among 4.00 to 7.20. Matching spots were 1025±24 in two repeats electrophoregrams and matching ratio was 96.30%. Conclusion The different protein spots were successfully established with high quality and sharpness separation by 2-DE and TOF-TOF-MS, which stands as a valuable resource for proteomics re-search of V. cholerae.
RESUMEN
Objective To develop a nested PCR for the detection of early syphilis and genotyping of Treponema pallidum (TP), and to investigate the distribution of genotypes of TP in Guangzhou. Methods Specimens were consecutively collected from genital ulcers of patients with suspected chancre during 2002-2004, and were detected by dark-field microscopy and nested PCR. The acidic repeat protein (arp) gene and the T. pallidum repeat (tpr) gene family were amplified with the positive specimens above. The number of repeats presented in the arp gene and the restriction fragment length polymorphism by Mse I in the tpr gene were analyzed by electrophoresis. The strains were genotyped according to Pillay's criteria. Results Out of 62 patients with suspected chancre, 33 cases (53.2%) were positive by dark-field microscopy and 54 cases (87.1%) by nested PCR. Of 47 TP-positive specimens genotyped by arp gene, 36 (76.6%) were type 14, while of 49 cases genotyped by tpr gene 39 (79.6%) were type d. By combining genotypes of arp and tpr genes, 7 genotypes were found, including 14d (31, 66.0%), 13d (5, 10.6%), 14b (4, 8.5%), 12b (3, 6.4%), 12d (2, 4.3%), 15d(l, 2.2%) and 14i (1, 2.2%). Conclusions Nested PCR shows a high sensitivity in early detection of TP. Genotype 14d seems the predominant type of TP in Guangzhou.