1.Establishment and research on Staphylococcus-infected-corneal C57BL/6 mouse model
Chun, LIU ; Zhiyun, YUAN ; Feifei, YANG ; Huaijin, GUAN ; Yixiang, SHAO
Chinese Ophthalmic Research 2010;28(3):199-202
Background An ethylnitrosourea(ENU)-induced mutant strain C57BL/6 mouse model has been established by our research group.This model is proved to have the spontaneous phenotype of corneal opacity and the typical pathological process similar to human keratitis.Therefore,this model is expected to be a good animal model in the research of the mechanism,hereditary property,and development of drugs for corneal infectious diseases.Objective The present study is to investigate the biological features of opportunistic pathogens using a mouse Staphylococcus-infected corneal model(C57BL/6 mouse) induced by N-ethyl-N-nitrosourea(ENU),and offers an evidence of stability in this animal model.Methods Ten-week-old male C57BL/6 mice were treated with ENU at 150mg/kg by intraperitoneal injection,and then mated with female mice after 60 days.Corneal opacity mutant mice in the F1 generation were selected to backcross with C57BL/6 mice.The bacteria were isolated from the eyeballs of the mutants and cultivated,purified and identified.Drug sensitivity assay was carried out to screen for effective antibiotics for clinic medical care.Results The staphylococcus-infected corneal mouse model(B6-Co) was established successfully,and the Staphylococcus sciuri strain was separated and purified,and then the sensitive antibiotics were distinguished from resistant ones.The sensitive drugs for Staphylococcus sciuri included azithromycin,clindamycin,chloramphenicol,gentamicin,rifampicin,tetracycline,amikacin,sulfamethoxazole compound sinomin,minocycline,levofloxacin,cephalothin,cefotaxime,and furazolidone;whereas this Staphylococcal strain was resistant to cefoxitin,penicillin,ampicillin,novobiocin.Nitrofurantoin showed an intermediate sensitivity.Conclusion The C57BL/6 mouse model is a spontaneous-derived animal model that is infected by coagulase-negative staphylococci,among which the most abundant strain is Staphylococcus sciuri.
2.Rapid detection of common bacterial infections of cerebrospinal fluid by genetics approach
Yixiang GUAN ; Jianhong SHEN ; Xingyun JU ; Demo WU ; Jinrong DING ; Yueping ZHONG ; Mingfei ZHANG ; Chunxiu ZHANG
Chinese Journal of Neurology 2012;45(8):586-589
ObjectiveTo assess gene chip application value in detecting pathogenic bacteria in intracranial infection cases.MethodsPrimers and probes aiming at the specific DNA sequences of 4 kinds of common pathogenic bacteria and 6 kinds of common drug resistance genes (DRGs) were designed and used to identify the bacteria and DRGs among 30 cerebrospinal fluid (CSF) specimens (12 positive,18negative in CSF culture) from patients with intracranial infection using multiplex polymerase chain reaction (mPCR) and gene chip.The results of gene detection were compared with those of CSF culture and drug sensitivity testing.ResultsBacteria were identified and DRGs were detected in 15 specimens; DRGs and 16S gene were detected in 8 specimens; neither bacterium nor DRG was detected in 7 specimens.ConclusionGene chip technique is characterized by its relative sensitivity and rapidity of detecting the pathogenic bacteria in CSF of intraeranial infection cases.