Designing And Using The Penicillium Marniffei Nucleate Probe

[Background and Objective]Penicilliosis marneffei(P.m),which is originated by Penicillium marniffei. Blue mold widely exist in the natural world,which confirm most of negative pathogenic bacteria in about 300 species blue mold,but Penicillium marniffei is the most pathogenicity in blue mold pathogenic bacteria. According to existing analysis of data, Penicilliosis marneffei is local epidemic,which is predilection at various kinds immune suppression patient. Now is a commom opportunistic pathogen seen in AIDS patients,our south country and Southeast Asia. In or of the past,diagnosing Penicillium marniffei usually depend on fungus cultivante,to result in delaying diagnosis. Penicillium marniffei easy mix up with histoplasmosis by pathology diagnosis.Thus,our research devote to design specificity Penicilliosis marneffei probe,in order to raising clinical diagnosis efficiency thoungh making use of in situ hybridization diagnosing Penicilliosis marneffei in Praffin tissue.[Method](1)As co-workers of the human genome research center of China in Shanghai,we have analysed the functional genes of P.m by methods of molecular biology and bioformatics.PYA92 was both constructed and analysed to find out specificity array. In NCBI, Penicilliosis marneffei analyze and compare with other Penicilliosis marneffei array ,and PYA92 compare with main pathogen in macrophage and fungus,finding out most conservatism gene,further designing the probe of in situ hybridization. We use PCR to amplification P.m definite part,and then depurating,sequencing these arrays,at last we use biotin labeling this conservation arrays.(2)Using this probe diagnose P.m and other deep mycosis in Paraffin tissue section,verification the specificity and the sensitivity of this probe.[Consequence]At patho-confirmed 23 cases of Penicilliosis marneffei tissue and 23 cases of other funguses(14 cases of Aspergillus,4 cases of Candida albicans,2 cases of cryptococcosis,3 cases of mold fungus ),which use in situ hybridization to diagnose,in order to verificate this probe .All of Penicilliosis marneffei tissue show masc,and its sensitivity is 100%;all of human gene and other funguses show negative ,and its specificity is 100%.thus ,we find out using in situ hybridization and pathology to diagnose can diagnosis coincidence in 100%. We initially verificate this probe and its actuality feasibility.[Conclusion](1)Using bioinformatics to analyze PYA92,in order to design the probe,we get the result that show this probe have hypersensitivity and specificity. we have necessity base in developing in situ hybridizat kit.(2)For subsequent research ,we have enlarge sample size and the kind of pathogen,advancing this technical stability and operating,in order to spread to basic hospital.