[Abstract] Objective To evaluate the effect of Mycobacterium tuberculosis Direct Assay (MTD) for rapid detecting Mycobacterium tuberculosis rRNA and Multi-locus PCR for M.bovis BCG strain typing in patients with suspected extra-pulmonary tuberculosis.Methods From June 2010 to December 2011,47 children and 75 adult patients with suspected extra-pulmonary tuberculosis in Shanghai public health clinical center were recruited.Also 48 non-tuberculosis patients were taken as a negative control.Clinical specimens from these patients were collected.Acid fast stain,solid culture,liquid culture,and MTD were used to detect all clinical specimens simultaneously.Screen tuberculosis strains of the culture isolates by MPT64 antigen assay and use Multi-locus PCR for the BCG strain genotyping of the isolates without MPT64 antigen.SPSS16.0 was used to analyse the results.Results The sensitivity for acid fast stain,solid culture,liquid culture and MTD test was 10.7％ (13/122),11.5％ (14/122),16.4％ (20/122) and 37.7％ (46/122),respectively.And the specificity of MTD was 100.0％.Six clinical isolates from children were identified as BCG by Multi-locus PCR typing,the same with chemical tests.Conclusions The MTD assay and the MGIT960 liquid culture are effective and reliable method for diagnosing extra-pulmonary tuberculosis.And Multi-locus PCR can be assisted for the early diagnosis of extra-pulmonary tuberculosis patients with suspected BCG infection.

Objective To investigate the correlation between the mutation of pncA gene and the susceptibility to pyrazinamide ( PZA) in Mycobacterium tuberculosis complex ( MTBC) strains and to analyze the mutation of panD and rpsA genes in wild type isolates without pncA gene mutation.Methods The sus-ceptibilities of 108 MTBC strains to first-line drugs including PZA were detected by using the MGIT 960 TB system.PCR was performed to amplify the 16S rDNA and pncA, panD and rpsA genes.The PCR products were analyzed by DNA sequencing analysis .Results Among the 78 multidrug-resistant MTBC strains , 47 isolates (60%) were resistant to PZA.Four out of 30 (13%) strains that were sensitive to ethambutol , iso-niazid, rifampicin and streptomycin (EIRS) were resistant to PZA.The drug-resistant MTBC strains showed higher resistance rate to PZA than that of the EIRS sensitive strains .There were 49 ( 96%) PZA-resistant isolates and 4 (7%) PZA-sensitive isolates occurred pncA gene mutation.Most of the pncA gene mutations in the genomes of PZA-resistant strains were base substitution mutation , especially the His57Asp substitu-tion.The pncA gene mutations centralized in the regions of 160-169, 203-289, 309-396 and 413-467.Seven novel mutation sites of pncA gene were observed including T175C, C188A, G insertion at 68, AGC insertion at 235, C insertion at 339, CC insertion at 392 and GT deletion at 395.The mutation sites founded in the genomes of PZA-sensitive strains were different from those of the PZA-resistant strains .No mutation of the pncA gene and the upstream regulatory sequence was found in two PZA-resistant strains , NJ44 and NJ108 . The sequence analysis of panD and rpsA gene showed that the NJ 108 strain had panD gene mutation at G419A, but no mutation was detected in the NJ 44 strain.Conclusion The multidrug-resistant MTBC strains showed higher resistance rate to PZA .The pncA gene mutation was common in PZA-resistant MTB strains and the panD gene mutation was also worthy of attention .

Abstract: Objective To investigate the willingness to receiving multitarget stool DNA (MT-sDNA) testing and factors affecting the payment among individuals receiving colonoscopy screening, so as to provide the evidence for the formulation and health economic evaluation of colorectal cancer screening strategies. Methods: Individuals at ages of 40 to 75 years that received colonoscopy screening in The Affiliated Hospital of Ningbo University Medical School from August 2021 to March 2022 were sampled. Participants' demographics, living behaviors, family history, willingness to receive MT-sDNA testing and willingness to pay for MT-sDNA testing were collected using questionnaire surveys, and factors affecting the willingness to receive and pay for MT-sDNA testing were analyzed using a multivariable logistic regression model. Results : A total of 546 respondents were enrolled, with a mean age of (56.25±8.66) years and including 282 men (51.65%). There were 504 respondents that were willing to receiving MT-sDNA testing (92.31%) and 480 that were willing to pay for the MT-sDNA testing (88.24%). Multivariable logistic regression analysis showed that a family history of colorectal cancer in first-degree relatives (OR=0.246, 95%CI: 0.068-0.888), history of hemorrhoids (OR=0.300, 95%CI: 0.109-0.826) resulted in low willingness to receive MT-sDNA testing, and recognizing the reliability of MT-sDNA testing (OR=5.749, 95%CI: 1.480-22.323), considering no difficulty in sampling for MT-sDNA testing (OR=32.042, 95%CI: 6.666-154.021) and considering a difficulty in sampling for MT-sDNA testing (OR=20.278, 95%CI: 4.405-93.354) resulted in high willingness to receive MT-sDNA testing, while recognizing the reliability of MT-sDNA testing (OR=5.003, 95%CI: 1.761-14.216), concern about the reliability of MT-sDNA testing (OR=4.166, 95%CI: 1.285-13.501), considering no difficulty in sampling for MT-sDNA testing (OR=6.558, 95%CI: 2.105-20.428) and considering a difficulty in sampling for MT-sDNA testing (OR=5.820, 95%CI: 1.810-18.720) resulted in high willingness to pay for the MT-sDNA testing among individuals receiving colonoscopy screening. Conclusion A family history of colorectal cancer in first-degree relatives, history of hemorrhoids and awareness of MT-sDNA testing are factors affecting the willingness to receive and pay for the MT-sDNA testing among individuals receiving colonoscopy screening.

ObjectiveTo analyze the characterstics of phenotype and genotype of multidrug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) by molecular line probe assay and liquid culture with MGIT960.MethodsGenoType MTBDR Kits were used for identifying the types of the first-line and second-line antituberculosis drug resistant genes partly and BD MGIT960 was used for detecting the chug susceptibility.Results( 1 ) Out of 94 MDR-TB strains,the rate of drug resistant to EMB,AMK,OFX and MFX by BD MGIT960 assay were 36.2%,17.0%,54.3% and 55.3%,respectively.Among these isolates,13 were extensively drug resistant tuberculosis (XDR-TB).(2) Compared with MGIT960,the concordance rate of GenoType MTBDRplus was 86.2% and 95.7% respectively.Taking MGIT960 results as reference,the sensitivity of GenoType MTBDRsl detecting the susceptibility of EMB,AMK,OFX and MFX to 94 isolates were 47.1%,81.3%,94.1%,94.2%,respectively.The specificity were 75.0%,98.7%,90.7%,92.9%,respectively.(3) Among the rpoB mutation categories,S531L accounts for most.MTB resistant to IFN caused by the mutation of katG chiefly and the S315T1 was in the majority.The gyrA mutation sites located at the ninety-fourth codon most.Out of 94 strains,23 were mixed with 2 kindsof Mycobacterium tuberculosis at least and 7 were undetectable mutations.Conclusion Among the M/XDR-TB,the strains resistant to INH,RFP,AMK,OFX and MFX were caused most by the mutation of katG,rpoB,rrs and gyrA,respectively.The relationship between EMB and embB was not so clear relatively.As a fast detecting drug susceptibility test kit,GenoType MTBDR possess good sensitivity and specificity.So,it could be as an assistant method to guide the therapy on clinic.

Objective:To improve the public welfare attributes of public hospitals through the optimization of the performance ap-praisal system of public hospitals,stimulate the enthusiasm of medical staff,and continuously improve the quality and level of medi-cal services.Methods:From the perspective of value co-creation,the three-party value subjects of hospitals,patients and depart-ments are considered,and the core indicators of Diagnosis Related Group(DRG)and Resource-based Relative Value Scale(RBRVS)accounting methods are introduced to construct a performance appraisal system at the level of medical departments in public hospi-tals.Results:The sample hospital applied DRG and RBRVS-based medical department performance appraisal system,and found that the equity of performance evaluation,the efficiency of hospital operation ability,and the satisfaction of patients and medical staff were significantly improved.Conclusion:Based on the theory of value co-creation,the performance appraisal system of the case hos-pital is optimized into three dimensions:cost control assessment,KPI index assessment and workload assessment,so as to achieve consistency between performance appraisal work objectives,and the the result variables of value co-creation,promote the balance of service quality and quantity of medical service providers,and promote the optimization of the effect of hospitals,departments and pa-tients.

Outbreaks of pseudorabies (PR) have occurred in southern China since late 2011, resulting in significant economic impacts on the swine industry. To identify the cause of PR outbreaks, especially among vaccinated pigs, 11 pseudorabies virus (PRV) field strains were isolated from Guangdong province during 2013–2014. Their major viral genes (gE, TK, gI, PK, gD, 11K, and 28K) were analyzed in this study. Insertions or deletions were observed in gD, gE, gI and PK genes compared with other PRV isolates from all over the world. Furthermore, sequence alignment showed that insertions in gD and gE were unique molecular characteristics of the new prevalent PRV strains in China. Phylogenetic analysis showed that our isolates were clustered in an independent branch together with other strains isolated from China in recent years, and that they showed a closer genetic relationship with earlier isolates from Asia. Our results suggest that these isolates are novel PRV variants with unique molecular signatures.
Asia ; China* ; Disease Outbreaks ; Genes, Viral ; Herpesvirus 1, Suid* ; Pseudorabies* ; Sequence Alignment ; Swine

Objective:To explore the serodiagnostic value of Rv0222/ESAT-6 fusion protein in tuberculosis.Methods:By using purified recombinant Rv0222/ESAT-6 fusion protein as antigen and enzyme-labeled antibody as secondary antibody,the serum of patients with tuberculosis and patients with human immunodeficiency virus(HIV)accompanied by tuberculosis,non-tubercu-losis patients and healthy people were detected with enzyme-linked immunosorbent assay(ELISA).The diagnostic value of the fusion protein in tuberculosis was evaluated.Results:By setting T lymphocyte spot test of infection with Mycobacterium tuber-culosis (T-SPOT)as the gold standard,the sensitivity and specificity of ELISA,of which Rv0222/ESAT-6 fusion protein was used as the antigen,in the diagnosis of tuberculosis were 86% and 100%,respectively.There was no statistically significant difference in the positive rate of tuberculosis between T-SPOT and ELISA.Conclusions:The ELISA with Rv0222/ESAT-6 fu-sion protein as antigen,has certain application value in the serodiagnosis of tuberculosis.

Objective To evaluate the performance of modified Hodge test on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Methods Fortynine Enterobacteriaceae isolates with decreased susceptibility to carbapenems ( MIC of imipenem, meropenem or ertapenem was ≥ 2 μg/ml ) were collected from 16 teaching hospitals from 2004 to 2008. MICs of imipenem, meropenem and etapenem were determined by agar dilution method. Carbapenemases were detected by modified Hodge test. Carbepenemase-causing positive results and AmpCs-causing positive results were differentiated by phenyl boronic acid and oxacillin. Beta-lactamases encoding genes including blaNDM-1were detected by PCR and sequencing. Results Thirty-six of 49 isolates were non-susceptible to imipenem (MIC >4 μg/ml), 31 were non-susceptible to meropenem (MIC > 4 μg/ml) and 47 were non-susceptible to ertapenem (MIC > 2 μg/ml). Twenty-three isolates showed positive modified Hodge test result, including 9 weak-positive results and 14 strong-positive results. Through PCR detection and sequencing, 2 out of 9 isolates showing weak-positive results carried blaKPC-2 and other 7 did not carry any carbapenemase genes but AmpCs/ESBLs genes. Among the 14 isolates showing strong-positive results, 4 carried blaKPC-2, 8 carried blaIMP-4 and 2 caried blaIMP-8. All 26 isolates with negative modified Hodge test result didn't carry any carbapenemase genes. No isolate carried blaNDM-1. Carbapenemases genes PCR detection was regarded as a gold standard, and the sensitivity, specificity, positive predictive value and negative predictive value of modified hodge test was 100%, 79%, 70% and 100% on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Conclusions Modified Hodge test revealed great sensitivity but showed a few false positive results. True and false positive results can be effectively differentiated by phynel boronic acid and oxacillin.