Objective To study the difference in detection rate of Mycobacterium tuberculosis in diffenent types of sumples.MethodsThe sputum,bronchial fluid,blood of 52 patients with clinical diagnosis of patients with pulmonary tuberculosis were tested with fluorescence quantitative PCR.Results45 cases were identiified in sputum and 51 cases were identified in bronehial perfusate and 15case were identified in blood.?2 was 4.875(P

Objective To develop an rapid, visuable method to detect the genotypes of HBV.Methods According to the published full-length sequence with known genotypes in GenBank, the specific primer and biotin-lablled probe were designed. We established a nucleic acid strip method for genotyping hepatitis B virus(HBV) isolates among 150 HBV infected patients and 20 healthy controls, and compared the results with those obtained by real-time PCR. Results There was 34.00% genotype B , 61.33% genotype C, 4.00% genotype B/C in 150 samples, respectively, while the remaining 0.67% unknown. The results for this assay were high comparable to the method of real-time PCR. Conclusion With the similar sensitivity and specifity when compared with real-time PCR, this rapid method is suitable to the clinical setting.

Objective To establish a rapid, sensitive, and specific quantitative method to detect hepatitis C virus. Methods A primer set targeting HCV 5'UTR was designed. The isothermal amplification was performed by the Bst DNA polymerase and AMV reverse transcriptase, under the temperature of 60℃ for 60 min. The signal was monitored by SYBR Green Ⅰ. Results One hundred and twenty positive serum samples, confirmed by the real-time PCR. All were detected by the isothermal amplification, while 110 healthy subjects' samples were negative by the both methods. The lower detect limit was determined to 10 IU/ml HCV-RNA, by the assay on serial dilutions of the quality control standards obtained from clinical investigation center of MOH. Conclusion A real time reverse loop-mediated isothermal amplification method was developed to detect HCV, with the characteristic of rapidity, high sensitivity and specificity.

Objective To investigate the distribution of hepatitis B virus genotypes in Hangzhou area and preliminarily identify and evaluate the applied characteristics of oligonuleotides(oligo)microarray for genotyping hepatitis B virus(HBV).Methods HBV PCR products were hybridized with oligonucleotide probes,which were prestablized on the chip.The hybridized results were colorized.According to the hybridization signal and the corresponding probe sequence,HBV genotype was determined.Results Of the 106 HBV DNA positive patients 37(34.91%)were genotype B,and 69(65.09%)were genotype C,No genotype A,E and F were found in the studied subjects.Conclusions HBV genotype B and C exsited in Hangzhou No HBV genotype A,D,E and F were found in the studied subjects.We should take further investigation for HBV genotypes by enlarging study population.The advantages of this assay are sensitive accurate,fast and economical when using it for HBV genotype test comparing with other relative methods.

Objective To investigate the distribution and virologic characteristics of HBV genotypes,and possible association with the severity of liver disease.Methods HBV genotype was determined,using oligonuleotides(oligo) microarray method.Results 37patients were differentiated as genotype B,69 genotype C.The clinical manifestations demonstrated that the serum lever of HBVDNA and HBeAg positive rate in patients of genotype C was 7.02?1.26 and 60.87% higher than 5.62?1.02and 32.43% in those of genotype B,the serum HBeAb positive rate in patients of genotype B was 67.57% higher than 39.13% in those of genotype C.The occurrence rates of those developed to chronic hepatitis B liver cirrhosis and hepatoma in those of genotype C were 46.38%,30.44% and 11.59% highter than that of 40.54%,13.51%and 8.11% in those of genotype B.Patients with genotype B were much younger than those with genotype C.Conclusions Genotype B and C exsited in Hangzhou,genotype C is a predominated genotype.The serum leverl of HBVDNA in those of genotype C is highter than that of genotype B.The positive rate of HBeAg in those of genotype B is highter than that of genotype C.The damage to liver induced by genotype C is severe than that of genotype B.

0.05). Conclusions E 3/3 and the prevalence of ? 3 allele were significantly higher in Shandong elderly population.

Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease (HFMD). Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovirus. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirns 71 and coxsackie virus AI6. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.

Objective To investigate the association of primary liver cancer(PLC)with the mutations of HBV precore and basic core promoter(BCP)genes.Methods The serum markers of hepatitis B and the quantities of serum HBV DNA were detected in 144 HBsAg-positive PLC patients.The precore and BCP gene mutations in patients with HBeAg-negtive and HBV DNA-positive were detected by real-time PCR.One hundred and twenty chronic hepatitis B(CHB)patients were randomly selected to serve as the conol.Results There were 46(3 1.94%)patients with HBeAg-positive and 98(68.06%)patients with HBeAg-negative.In 98 HBeAg-negative patients,56(57.14%)were HBV DNA-positive,in which 43 (76.79%)were with precore 1896 gene mutations,50(89.29%)were with BCP1762/1764 gene mutations.and 38(67.86%)were with both gene mutations.Precore 1896 and BCP1762/1764 gene mutation rates in PLC patients were much higher than those in CHB patients(χ2=9.36 and 5.77,P＜0.05).Conclusion PLC may be associated with the mutations of HBV precore anti BCP genes.

OBJECTIVE To investigate the current status,the clinical features and the pathogens of invasive fungal infections in hospital in order to provide clinical treatment based on identification and susceptibility test.METHODS The fungus-cultured positive cases among the discharged patients from Jan 2004 to Nov 2006,were analyzed according to their definite diagnosis of invasive fungal infections under the items,such as the patients age,underlying disease,sample,strain,and species distribution.RESULTS The rates of invasive fungal infections were 4.26%.There were 2221 fungus strains belonged to 8 species in all samples;the patients age was 7-96 years with 2 kinds of various underlying diseases;the age of 2221 cases was 60 years old,mainly senile patients with various diseases accounted for 68.29%.Lower respiratory tract was the most frequent infection site.The main pathogens of invasive fungal infections were Candida spp(93.38%).Strains of Candida albicans were the most frequent organism isolated accounted for 66.19% of all the isolates.C.glabrata,C.krusei and C.tropicalis accounted for 9.19%,8.10% and 4.50%,respectively,the others accounted for only 6.32%.The main infected sites were lower respirtory tract,urinary tract and digestive tract.CONCLUSIONS Candida spp are still the main pathogens of invasive fungal infections.The epidemiological properties of invasive fungal infections is changed.The incidence of non-C.albicans and the Aspergillus strains that arouse invasive infections is increasing recently.

Objective To investigate the distribution of hepatitis B virus(HBV)genotypes in Zhejiang.Methods HBV genotyps were detected by fluorimetric real-time PCR in 240 HBVDNA positive patients who were born in Zhejiang(Hangzhou,Huzhou,Jinhua,Shaoxing,Taizhou,Ningbo each 40 positive patients).Results of the 240 HNBDNA positive patients,82(34.17%)were genotype B,and 140(58.33%)were genotype C,15(6.25%)were genotype B and C,3(1.25%)were genotype D.No genotype A、E and F found in the studied subjects.Conclusions HBV genotype B,genotype C,genotype B and C,genotype D existed in Zhejiang and there is no difference in ferms of the distribution of genotypes in six areas metioned above.