Explore For The Existence Of Nature Chlamydia Trachomatis Phage

C.T lives an unique intracellular parasitism life,grow within a specialized cytoplasmic compartment know as an "inclusion" and grows by binary fission within the inclusion. C.T mainly lives on the epithelial cell with human as it natural host. It leads to many diseases. Genitourinary tract infections caused by C.T has become the most common STD in recent years. Its recurrency and peresistentency and had no effective vaccine make it to the the community public health problem.The research on CT infection is the hot spot and focal point at home and abroad.It is necessary to take some strong prevention and control measures to improve the popular of the status of C.T infection. C.T phage was recognized as the most promising and effective means to solve this problem, but it has not been found yet.Chlamydia phage is an invasive virus of chlamydia,.It could lead to the disintegration of Chlamydia. It belongs to Chlamydiamicrovirus of Microviridae of group Ⅱ-single-stranded DNA virus. So far, we have found Chlamydia phage Chpl, Chp2, Chp3, Chp4phiCPAR39and phiCPGl. But, there is no report about the phage of Chlamydia trachomatis yet. According to the phylogenetic tree,C.T and C. psittaci are more acient Chlamydias than C. pneumonia with more complex structure and many types.Whererase, C. pneumonia has its phage already and the same to C. psittaci, there shoulde be C.T phage.If,we can find it luckily,the potential value of clinical is amazing.Phage adsorbed on outer membrane of sensitive chlamydial depends on the combination between the receptor protein and phage capsid protein Vp123. Where Vpl is the major with the largest molecular weight and is highly conserved and specific,can be taken to a good marker to search for C.T phage.Objective:Collect a variety of specimens in different clinical conditions.Screening for Vpl gene and antibody against Vpl protein of chlamydia trachomatis phage from clinical secretions and serums. Trying to find the existence of nature C.T phage and lay the foundation for the resolution of C.T infections.Methods:We collected swabs and serums from STD clinic patients. We designed capsid protein Vpl gene primers of C.T phage, used PCR to detect it. The susscessful coloned Vp1gene products were send to sequence.Meanwhile,we cultured the positive swabs in McCoy cells normally or with sereotype E strain together.post the infection of72hs,detect the Vpl protein of Chlamydia phage by immunofluorescence. With ELISA and Western Blot to find out antibody against Vpl protein of Chlamydia phiCPGl in serum.Results:Screen36positive swabs from1542swabs.in Vpl gene, including859cases of C.T (+) patients and683cases of C.T (-) patients; Sequencing results showed that:There are only4to5bases difference between amplified Vpl gene sequencesand the target fragment.Blast the sequences on the NCBI, coverage and similarity are both more than90%with Chlamydia phage phiCPGl, chp4, CPAR39, chp3, chp2. But clinical isolates screened by immunofluorescence got no positive results. Obtained6positive serum results consist in5from267cases of C.T (+) patients and1from186cases of C.T (-) patients;Conclusions:From clinical swabs and serums we successfully found out Vpl gene and Vp1protein of chlamydial trachomatis phage. Further studies are needed to confirm it.