Applications of molecular diagnostics in clinical microbiology laboratory were introduced in the article,including nucleic acid hybridization,nucleic acid amplification,DNA sequencing,gene chips and mass spectrometry.Molecular diagnostic techniques provide major tools for rapid diagnosis of infectious diseases,molecular epidemiology investigation,rapid identification of microbial pathogens and the study of the pathogenicity and antibiotics resistance.The application of these new technologies,as the complement of traditional culture methods,increases sensitivity,accuracy and diagnostic efficiency of assays.

Recent major changes and updates of Clinical and Laboratory Standards Institute(CLSI) document M100 for performance standards for antimicrobial susceptibility testing were introduced in the article,which include changes of interpretive criteria and comments,antimicrobial susceptibility testing of infrequently isolated or fastidious bacteria,changes of appendixes and others,Brief comments were made for these changes.

In recent years , respiratory tract infectious diseases and public health events caused by human adenovirus type 7(HADV-7) had been rising.Especially in Asia, the outbreaks of acute respiratory illness caused by HADV-7, accounting for more than 60%in the world, had brought great influence to the public health.In order to prevent the occurrence of HADV-7 epidemic and improve the diagnostic performance of laboratory , the research progress of HADV biological features and typing , HADV-7 epidemiological characteristics , clinical and differential diagnosis and laboratory examination methodologies were reviewed in this paper.

Major changes and updates of Clinical and Laboratory Standards Institute (CLSI) document M100-S22 for performance standards for antimicrobial susceptibility testing were introduced in the article,which includes ( 1 ) general changes; (2) changes of drugs recommended for testing and reporting;( 3 ) changes of interpretive criteria ( breakpoints ) and comments ; ( 4 ) changes of quality control and others; and (5) changes of appendixes and glossaries.

0.05). VIP, however, could inhibit the Con A-induced proliferation of T-cells from control subjects more significantly than that from asthmatics (P0.05). The cAMP level in T-cells, however, increased more significantly in the control group than that in the asthmatic group after the treatment of VIP or NaF (P0.05). CONCLUSION: Inhibition effect of VIP on Con A-induced proliferation of T-cells was less in asthmatics than in control subjects, which may be related to insufficiency of Gs ? coupled VIP receptor on T-lymphocytes in asthmatics.

Objective To explore the method of Oncomelania hupensis snail surveillance in transmission?interrupted re?gions of schistosomiasis,so as to provide a new pattern for the surveillance of snail status in the transmission?interrupted areas. Methods In Dongtai County,north of Jiangsu Province where the transmission of schistosomiasis was interrupted,the surveil?lance of snail status was performed by means of the village?level general survey,village?level sampling survey,snail survey in key settings of township and county,and snail reporting by residents. In addition,quality?control snail sites were set up for quali?ty control. The effectiveness and cost of the four methods of snail surveillance was compared. Results A total of 163 079 set?tings and 22 785.62 hm2 were surveyed in Dongtai County from 2008 to 2013,and 89 residual snail breeding sites were found, with a snail area of 1.48 hm2. The patterns of village?level general survey,village?level sampling survey,and snail survey in key settings of township and county were employed for investigation of 94 550,45 033 and 23 496 settings,consisting of 57.98%, 27.61%and 14.41%of total settings,and 2,0 and 6 settings were found with snails,with 0.021/thousand,0,and 0.255/thou?sand detection rates of settings with snail breading sites,respectively. The pattern of snail survey in key settings of township and county was more effective than that of village?level survey to detect snail breeding sites(χ2 =19.158,P=0). The recovery rate of quality?control snail breeding sites was 52.56%,38.27%and 73.62%for the three patterns of snail survey,respectively,and the pattern of snail survey in key settings of township and county showed a higher quality for snail survey than that of village?lev?el survey(χ2 =111.597 and 85.991,both P=0). During the period from 2008 to 2013,289 person?times reported 279 sus?pected snail habitats and 1 501 living snails,and no Oncomelania snails were found. During the 6?year period,there were 1617.5,964.7 and 527.7 thousand RMB invested for village?level general survey,village?level sampling survey,and snail sur?vey in key settings of township and county,and the cost of snail survey per hm2 was 129.88,133.6 and 162.57 RMB/hm2,re?spectively. The cost of village?level general survey and snail survey in key settings of township and county to detect a snail?breed?ing site was 808 800 and 88 000 RMB,respectively(t=12.850,P=0.000),and the cost of snail survey in key settings of town?ship and county was 10.88%of that of village?level general survey. Conclusion The snail survey in key settings of township and county is a highly effective and high?quality method for snail surveillance,which may serve as a prior way for the surveil?lance of snail status in the transmission?interrupted areas of schistosomiasis.

Objective To investigate the molecular characteristics and epidemiological signification of patients with low-level HBsAg. Methods PCR and gene sequencing were used to detect HBV DNA and Tyr-Met-Asp-Asp(YMDD) mutant in 136 serum samples with low-level HBsAg and 44 sernm samples with high-level HBsAg. Genotyping was performed in 47 cases with HBV DNA 10~5 copies/L by concentration method and 37 cases with high-level HBsAg. S gene sequences and serotypes were analyzed in 14 cases with HBV DNA 105 copies/L and 29 cases with high-level HBsAg. S gene sequences were compared with the consensus sequence of Chinese strain by BioEdit software. Results The HBV DNA-positive rate, YMDD mutation rate and HBV DNA load (logarithm) in low-level and high-level HBsAg group were 34.6% (47/136), 0% (0/136), 6.5±1.4 and 84.1% (37/44), 9.1% (4/44), 8.9±1.8, respectively. There was statistically significant differences between two groups (for concentration method,χ~2 = 30.8, P < 0.05; for direct method, χ~2 = 53.5, P < 0.05; for YMDD mutation ratio, P = 0.003, For HBV DNA (log), t = 6.5, P < 0.05). The genotypes in low-level HBsAg group included type B (16/47), type C (5/47) and non-classified ones(26/47). There were significant differences between two groups (χ~2=21.8, P <0.01). The serotypss included adw (7/14), ayw (4/14), adr (2/14) and ayr (1/14). There were significant differences in genotypes (χ~2 = 13.5, P < 0.05) but not in serotypes between two groups (χ~2 = 4.7, P >0.05). S gene sequencing results showed no S gnne variation was detected, but there were 6 single nucleotide polymorphisms in 16 cases, which would not result in the alternation of amino acid. Conclusions Low-replication phenomenon of HBV DNA was present in patients with low-level HBsAg. The major genotyps and serotype was type B and adw/ayw, respectively. Polymorphic variants have been found in the S gene. The existence of low-level HBsAg might be related with its own molecular characteristics resulting in low expression of HBsAg or immune tolerance induced by low-level HBsAg after HBV infection.

Objective To evaluate the performance of Carba NP test (CNP) for detection of carbapenemases in carbapenem-non-susceptive Enterobacteriaceae.Methods Carbapenemases in Enterobacteriaceae strains isolated from clinical specimens in the 117th hospital of PLA from January 2010 to December 2014 were detected by both CNP and modified Hodge test (MHT),and corresponding results were analyzed and compared by Chi-square using SPSS version 19.0.Data acquired from PCR were used as the standard.Results Of the 253 carbapenem-non-susceptive Enterobacteriaceae strains,188 positive strains and 65 negative strains were detected by MHT,and 175 positive strains,77 negative strains and 1 undetectable strain were detected by CNP,while 177 positive strains and 76 negative strains were detected by PCR.Sensitivities of two tests were comparable [CNP,98.9％ (175/177),versus MHT,98.3％ (174/177);x2 =0.5,P =0.48],but CNP was more specific [100.0％ (76/76) versus 81.6％ (62/76);x2 =12.1,P ＜ 0.05].Positive result in the CNP was emerged earliest in a KPC-producing strain.Conclusion The CNP is rapid and simple,which have good sensitivity and specificity.Yet further perfection is still needed in the future.

Objective To discuss the risk factors for susceptibility of subcutaneous tophi with an aim to provide clinical evidence for the prevention and treatment of tophi.Methods A total of 5 321 cases of gout patients whose course of disease was less than 10 years were selected and divided into two groups according to whether a subcutaneous tophus was present. The clinical information was collected and relevant biochemical indices were detected.Results There were significant differences in the ratios of regular exercise, involvement of upper limb joints, combining renal insufficiency, kidney stone and coronary heart disease between the tophus group and the non-tophus group(P<0.05 or P<0.01). There were significant statistical differences in age, disease duration, body mass index, systolic blood pressure, diastolic blood pressure, serum uric acid, serum creatinine, creatinine clearance rate, the number of joints involved between the two groups[(51.80±14.58 vs 48.45±14.45)year,(5.61±3.20 vs 3.28±3.06)year,(26.50±3.39 vs 27.05±3.45)kg/m2,(138±20 vs 133±18)mmHg, 1 mmHg=0.133 kPa,(89±13 vs 87±12)mmHg,(486.66±121.96 vs 460.02±119.10) μmol/L,(112.11±62.97 vs 104.43±46.28) μmol/L, (86.46±37.99 vs 93.00±40.11 )ml/min,(2.89±1.86 vs 1.76±1.22) joints, all P<0.01]. There were no statistical differences in family history, gender, history of smoking and drinking, the means of waistline, fasting blood glucose, triglyceride, and total cholesterol between the two groups(all P>0.05). Further logistic regression analysis indicated that disease duration, a large number of joints involved, the involvement of upper limb joints, combining kidney stones, higher serum uric acid level, and higher diastolic pressure were risk factors of the early onset of subcutaneous tophi, while regular exercise and body mass index were protective factors.Conclusion Patients with long duration, high serum level of uric acid, a large number of joints involved, upper limb joint involved, kidney stones, and hypertension were more likely to develop subcutaneous tophi. These risk factors should be intervened actively in clinic.

OBJECTIVE To investigate the distribution and the drug resistance of Klebsiella pneumoniae in our hospital,and provide reference for the rational use of antimicrobial agents in clinic.METHODS The strains of K.pneumoniae isolated from clinical samples were identified by the VITEK-32 system.Extended spectrum ?-lactamases(ESBLs) were determined by phenotypic confirmatory test.Susceptibility test to antimicrobial agents was performed by disk diffusion methods and analyzed by WHONET 5.3.RESULTS 93.6% Of total 256 strains of K.pneumoniae were isolated from urine,sputum,blood and secretion samples,and 54.3% of them were isolated from sputum.81.6% Of infections caused by K.pneumoniae were frequently occurred in patients in intensive care units(ICU),departments of respiratory medicine and gastroenterology,departments of hematology and endocrinology,cadred wards and department of nephrology.The isolated ratio of ESBLs producing K.pneumoniae were 39.8%.The rates of resistance to piperacillin,cefuroxime,ceftazidime, cefotaxime,ceftriaxone,aztreonam,gentamicin,ciprofloxacin,and sulfamethoxazole/trimethoprim were between 32.4% and 50% in all strains;and the resistance rate to cefoperazone/sulbactam,piperacillin/tazobactam,cefepime,cefoxitin,and amikacin was less than 22.7%.There were isolates producing ESBLs,which showed much higher resistance to antimicrobial agents tested(but susceptible to imipenem and meropenem) than that of non-ESBLs-producing K. pneumoniae(P