| Gonorrhea is a worldwide sexually transmitted disease (STD). In our country the number of gonococcal infectious patients is increasing rapidly in the last 20 years. Because of extensive transmission, gonorrhea seriously endangers mental and physical health of the mankind and social stability as well. Until now, we can't effectively control gonococcal infections, which is probably due to the deficiency of convenient and rapid laboratory test Therefore, to develop a simple and practical laboratory test would be benefit to abolishing pathogen and controlling the disease spread.The methods of current gonococcal laboratory examination include microscopic smear, bacterial culture, leukocyte esterase test (LET), antigen detection and nucleic acid (NA) assay etc. But the detection of gonococcal infection is mainly depending on microscopic smear and bacterial culture. Of all current laboratory tests, every method has limitation during practical application. Misdiagnosis and false negative easily occurs by microscopic smear, particularly in female. Although the method of bacterial culture is a "gold standard ", its culture requirements are very fastidious. It is not simple and convenient, and the resistance of gonococci to physical and chemicalfactor is weak. The result needs to be waited for more than two days and it is not suit for screening low-risk population. LET isn't as specific as people wished. The antigen examination hasn't been recommended to be employed as a routine gonococcal diagnosis method. The nucleic acid assay has strict requirements to experiment conditions, and false positive result can't be gotten rid of. Taken together, of all gonococcal detections, traditional bacterial culture is not convenient and is time costing. All other non-culture methods are not ideal enough either. Therefore, no specific, rapid, simple and low cost diagnostic kit is available for detecting gonococci, especially in low-risk of the population of gonococcal infections.Theoretically, the antigen detection can be applied in bacterial infectious disease, for example gonorrhea, TB etc. But many major surface antigens (such as pili, PII, PHI and LPS) of NG undergo antigenic variation or antigen loss during natural infection, so that gonococci can evade immune attacking, and it makes the antigen detection no longer effective. But many researches show gonococcal outer membrane protein PI does not undergo antigenic variation during natural infection. Porin is the major protein on the surface of Neisseria gonorrhoeae (NG). Gonococcal isolates are divided into serotypes on the basis of porin immunoreactivity .The two main forms PIA and PIB are further subdivided into serovars defined by reactions with panels of monoclonal antibody. PI may play an important role in pathogenesis, by virtue of its ability to interact directly with the cell membrane of eukaryotic cells. PI is also a major target for immune attack on the gonococcus. A lot of data show PI has the virtue of stability and antigenicity. But the research on PI by cloning, expression and purification with GST system is still very few, and its immunogenic character in animals still needs to be further researched.8In order to investigate the immunogenic character of self-cloned protein PI, in this experiment we prepared rabbit polyclonal antibodies to GST-PI protein by immunising rabbit with our self-produced protein. We hope to find a new simple and practical laboratory examination method, which would be hopefully benefit to controlling gonococci infection.Material and Method Animal strainsZhejiang domestic rabbits: all three domestic rabbits were male and about 2.5 Kg of wL ClinictI dataThe number of gonorrhea patients enrolled is 35, including 29 adult male with ages ranging from 22 to 54; 5 adult female with ages ranging from 18 to 58; 1 child with 5 years old. All patients of gonorrhea came from Department of Dermatology and Venerology of Sir Run Run Shaw Hospital and the Second Hospital attached to Medical School of Zhejiang University. |
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