Quantitative microbiological risk assessment of nontyphoidal Salmonella in ground pork in households in Chengdu, China.

Foodborne disease caused by nontyphoidal Salmonella (NTS) is one of the most important food safety issues worldwide. The objectives of this study were to carry out microbial monitoring on the prevalence of NTS in commercial ground pork, investigate consumption patterns, and conduct a quantitative microbiological risk assessment (QMRA) that considers cross-contamination to determine the risk caused by consuming ground pork and ready-to-eat food contaminated during food handling in the kitchen in Chengdu, China. The food pathway of ground pork was simplified and assumed to be several units according to the actual situation and our survey data, which were collected from our research or references and substituted into the QMRA model for simulation. The results showed that the prevalence of NTS in ground pork purchased in Chengdu was 69.64% (95% confidence interval [CI], 60.2-78.0), with a mean contamination level of -0.164 log CFU/g. After general cooking, NTS in ground pork could be eliminated (contamination level of zero). The estimated probability of causing salmonellosis per day was 9.43E-06 (95% CI: 8.82E-06-1.00E-05), while the estimated salmonellosis cases per million people per year were 3442 (95% CI: 3218-3666). According to the sensitivity analysis, the occurrence of cross-contamination was the most important factor affecting the probability of salmonellosis. To reduce the risk of salmonellosis caused by NTS through ground pork consumption, reasonable hygiene prevention and control measures should be adopted during food preparation to reduce cross-contamination. This study provides valuable information for household cooking and food safety management in China.

Characterization of Non-O157 Shiga Toxin-Producing Escherichia coli Cultured from Cattle Farms in Xinjiang Uygur Autonomous Region, China, During 2016-2017.

Most outbreaks of Shiga toxin-producing Escherichia coli (STEC) are attributed to consumption of contaminated foodstuffs including beef and dairy products. In this study, we evaluated the prevalence of non-O157 STEC cultured from beef and dairy cattle and collected in Xinjiang Uygur Autonomous Region in China. Results identified 67 non-O157 STEC recovered from the 793 samples including beef cattle (10.28%, 43/418) and dairy cattle (6.40%, 24/375). A total of 67 non-O157 STEC was sequenced allowing for in silico analyses of their serotypes, virulence genes, and identification of the corresponding multilocus sequence types (STs). Twenty-one O serogroups and nine H serotypes were identified and the dominant serotype identified was O22:H8. One stx1 subtype (stx1a) and four stx2 subtypes (2a, 2b, 2c, and 2d) were found in the 67 non-O157 STEC isolates. The results revealed that stx1a+stx2a-positive STEC isolates were predominant (32.83%, 22/67), followed by stx1a+stx2d (29.85%, 20/67) and stx2a alone (17.91%, 12/67). Non-O157 STEC isolates carried virulence genes ehxA (98.51%), subA (53.73%), and cdtB (17.91%). Of the four adherence-associated genes tested, eaeA was absent, whereas lpfA and iha were present in 67 and 55 non-O157 STEC isolates, respectively. The STEC isolates were divided into 48 pulsed-field gel electrophoresis patterns and 10 STs, and ST446 (O22:H8) was the dominant clone (22.38%). Our results revealed that there was a high genetic diversity among non-O157 STEC isolated from beef and dairy cattle, some of which have potential to cause human diseases.

The Epidemiology of Monophasic Salmonella Typhimurium.

Salmonella enterica remains an important foodborne pathogen in all regions of the world, with Typhimurium as one of the most frequent serotypes causing foodborne disease. However, the past two decades have seen a rapid worldwide emergence of a new Salmonella serotype, namely monophasic variant of S. Typhimurium, whose antigenic formula is 1,4,[5],12:i:-. It has become one of the 2-5 most common Salmonella serotypes responsible for animal and human infections in different regions. The global epidemic of monophasic S. 1,4,[5],12:i:- has mainly been characterized by an increase in multidrug-resistant S. 1,4,[5],12:i:- isolated in Europe since 1997. The unexpected link to swine has escalated monophasic S. Typhimurium infections to the status of a global public health emergency. The large-scale application of whole genome sequencing (WGS) in the last 10 years has revealed the phylogenetic associations of the bacterium and its antimicrobial resistance (AMR) genes. Local and global transmission reconstructed by WGS have shown that different clones have emerged following multiple independent events worldwide, and have elucidated the role of this zoonotic pathogen in the spread of AMR. This article discusses our current knowledge of the global ecology, epidemiology, transmission, bacterial adaptation, and evolution of this emerging Salmonella serotype.

Detection of plasmid-mediated tigecycline-resistant gene tet(X4) in Escherichia coli from pork, Sichuan and Shandong Provinces, China, February 2019.

The plasmid-mediated high-level tigecycline resistance gene, tet(X4), was detected in seven Escherichia coli isolates from pork in two Chinese provinces. Two isolates belonged to the epidemic spreading sequence type ST101. Tet(X4) was adjacent to ISVsa3 and concurrent with floR in all seven isolates. In addition to IncFIB, the replicon IncFII was found to be linked to tet(X4). This report follows a recent detection of tet(X3)/(X4) in E. coli from animals and humans in China.

Soil microbial communities of three major Chinese truffles in southwest China.

Tuber pseudoexcavatum, Tuber sinoaestivum, and Tuber indicum are the 3 most important truffles growing in southeast China; however, their cultivation is still inefficient owing to the lack of understanding regarding the composition and function of the bacterial and fungal communities from the soils around the fruit bodies and the ectomycorrhiza of these truffles. The aim of this study was to disclose the microbial communities in truffle-producing soils in Huidong County, Sichuan, China, by using barcoded pyrosequencing. Approximately 350 000 quality-controlled sequences were obtained and grouped into 14 025 bacterial operational taxonomic units (OTUs) and 4385 fungal OTUs, which included 29 bacterial and 7 fungal phyla, respectively. The bacterial genus Acidobacterium and fungal genera Modicella, Pseudogymnoascus, and Mortierella were significantly more abundant in the control soils than in the truffle-producing soils (P < 0.05), while the bacterial genus Sphingomonas (Alphaproteobacteria) and arbuscular mycorrhizal fungal genus Glomus were significantly enriched in truffle-producing soil than in the control (P < 0.05), indicating their different roles within truffle grounds. Notably, some nonfungal organisms detected by 18S rDNA pyrosequencing were of high abundance, among which Cercozoa and Ochrophyta were significantly (P < 0.05) more abundant in truffle soils than in control soils, indicating their interactions with truffles.

Mixed Infections and Rifampin Heteroresistance among Mycobacterium tuberculosis Clinical Isolates.

Mixed infections and heteroresistance of Mycobacterium tuberculosis contribute to the difficulty of diagnosis, treatment, and control of tuberculosis. However, there is still no proper solution for these issues. This study aimed to investigate the potential relationship between mixed infections and heteroresistance and to determine the high-risk groups related to these factors. A total of 499 resistant and susceptible isolates were subjected to spoligotyping and 24-locus variable-number tandem repeat methods to analyze their genotypic lineages and the occurrence of mixed infections. Two hundred ninety-two randomly selected isolates were sequenced on their rpoB gene to examine mutations and heteroresistance. The results showed that 12 patients had mixed infections, and the corresponding isolates belonged to Manu2 (n = 8), Beijing (n = 2), T (n = 1), and unknown (n = 1) lineages. Manu2 was found to be significantly associated with mixed infections (odds ratio, 47.72; confidence interval, 9.68 to 235.23; P < 0.01). Four isolates (1.37%) were confirmed to be heteroresistant, which was caused by mixed infections in three (75%) isolates; these belonged to Manu2. Additionally, 3.8% of the rifampin-resistant isolates showing no mutation in the rpoB gene were significantly associated with mixed infections (χ(2), 56.78; P < 0.01). This study revealed for the first time that Manu2 was the predominant group in the cases of mixed infections, and this might be the main reason for heteroresistance and a possible mechanism for isolates without any mutation in the rpoB gene to become rifampin resistant. Further studies should focus on this lineage to clarify its relevance to mixed infections.

Association of CRISPR/Cas evolution with Vibrio parahaemolyticus virulence factors and genotypes.

Clustered regularly interspaced short palindromic repeats (CRISPR), which is considered to be an immune system for bacteria, has been widely used as a tool for genome editing and genotyping. It has also been reported to be associated with virulence factors in some bacteria. To understand the role of CRISPR in the virulence and evolution of pathogenic Vibrio parahaemolyticus, 154 V. parahaemolyticus strains isolated from clinical samples and 54 strains from food samples taken in Shenzhen, China were subjected to a correlation analysis of CRISPR and virulence factors TDH and TRH. We also performed multilocus sequence typing (MLST) for genotype analysis. Six different CRISPR sequence types (CSTs) of V. parahaemolyticus were identified, and CSTs were found to be significantly associated with the virulence factors tested and MLST genotype. Therefore, CSTs provide insight into the evolution of V. parahaemolyticus. Moreover, identification of CSTs may lend insight into the virulence potential of strains.

Vibrio parahaemolyticus, Southern Coastal Region of China, 2007-2012.

We analyzed the prevalence and characteristics of Vibrio parahaemolyticus among patients with acute infectious diarrhea in the southern coastal region of China. V. parahaemolyticus was the leading cause of bacterial infectious diarrhea in this region during 2007-2012. Serotype O3:K6 strains were most common, followed by serotypes O4:K8 and O3:K29.

Prevalence and Genomic Characteristics of mcr-Positive Escherichia coli Strains Isolated from Humans, Pigs, and Foods in China.

Colistin is one of the last-resort antibiotics for treating infections caused by multidrug-resistant (MDR) Gram-negative bacteria. However, mcr genes conferring resistance to colistin have been widely identified, which is considered a global threat to public health. Here, we investigated the prevalence and characteristics of mcr-harboring Escherichia coli strains isolated from humans, animals, and foods in China by PCR, antimicrobial susceptibility testing, conjugation experiments, molecular typing, genome sequencing, and bioinformatics analysis. In total, 135 mcr-1-harboring E. coli isolates were acquired from 847 samples, and 6 isolates carried mcr-3. Among them, 131 isolates were MDR bacteria. Sixty-five resistance genes conferring resistance to multiple antimicrobials were identified in 135 isolates. The diverse pulsed-field gel electrophoresis (PFGE) patterns and sequence types (STs) of mcr-1-carrying isolates demonstrated that clonal dissemination was not the dominant mode of mcr-1 transmission. Seven types of plasmids were able to carry mcr-1 in this study, including IncI2, IncX4, IncHI2, p0111, IncY, and two hybrid plasmids. The genetic structures carrying mcr-1 of 60 isolates were successfully transferred into the recipient, including 25 IncI2 plasmids, 23 IncX4 plasmids, and an IncHI2 plasmid. mcr-1-pap2 was the dominant mcr-1-bearing structure, followed by ISApl1-mcr-1-pap2-ISApl1 (Tn6330) and ISApl1-mcr-1-pap2, among 7 mcr-1-bearing structures of 135 isolates. In conclusion, IncI2, IncX4, and IncHI2 plasmids were the major vectors spreading mcr-1 from different geographical locations and sources. The prevalence of Tn6330 may accelerate the transmission of mcr-1. Continuous surveillance of mcr-1 and variants in bacteria is vital for evaluating the public health risk posed by mcr genes. IMPORTANCE The spread of polymyxin-resistant Enterobacteriaceae poses a significant threat to public health and challenges the therapeutic options for treating infections on a global level. In this study, mcr-1-bearing ST10 E. coli was isolated from pigs, pork, and humans simultaneously, which demonstrated that ST10 E. coli was an important vehicle for the spread of mcr-1 among animals, foods, and humans. The high prevalence of mcr-1-positive E. coli strains in pigs and pork and the horizontal transmission of mcr-1-bearing plasmids in diverse E. coli strains suggest that pigs and pork are important sources of mcr-1-positive strains in humans and pose a potential threat to public health. Additional research on the prevalence and characteristics of mcr-1-positive E. coli is still required to facilitate early warning to improve polymyxin management in hospitals.

Identification of Mobile Colistin Resistance Gene mcr-10 in Disinfectant and Antibiotic Resistant Escherichia coli from Disinfected Tableware.

The widespread escalation of bacterial resistance threatens the safety of the food chain. To investigate the resistance characteristics of E. coli strains isolated from disinfected tableware against both disinfectants and antibiotics, 311 disinfected tableware samples, including 54 chopsticks, 32 dinner plates, 61 bowls, 11 cups, and three spoons were collected in Chengdu, Sichuan Province, China to screen for disinfectant- (benzalkonium chloride and cetylpyridinium chloride) and tigecycline-resistant isolates, which were then subjected to antimicrobial susceptibility testing and whole genome sequencing (WGS). The coliform-positive detection rate was 51.8% (161/311) and among 161 coliform-positive samples, eight E. coli strains were multidrug-resistant to benzalkonium chloride, cetylpyridinium chloride, ampicillin, and tigecycline. Notably, a recently described mobile colistin resistance gene mcr-10 present on the novel IncFIB-type plasmid of E. coli EC2641 screened was able to successfully transform the resistance. Global phylogenetic analysis revealed E. coli EC2641 clustered together with two clinically disinfectant- and colistin-multidrug-resistant E. coli strains from the US. This is the first report of mcr-10-bearing E. coli detected in disinfected tableware, suggesting that continuous monitoring of resistance genes in the catering industry is essential to understand and respond to the transmission of antibiotic resistance genes from the environment and food to humans and clinics.

Urban resilience assessment based on contradiction between supply and demand of the daily activity-environment system: A case study on Nanjing, China.

In addition to safety and risk issues, urban resilience research should focus more comprehensive and universal contradictions. Considering the main contradictions of Chinese urban development, and the connotation of urban resilience, a theoretical framework is constructed based on the contradiction between supply and demand of the daily activity-environment system. Furthermore, an assessment index system of urban resilience is also developed that considers both volume and quality of demand and supply and has a grade standard. Finally, utilizing multi-source geographic big data, Nanjing is taken as a case study, the spatial characteristics and optimization path of urban resilience are identified. The main conclusions are as follows: (1) The activity-environment system resilience presents a "center-periphery" sprawling spatial structure, and it is mainly at the level of "reluctant resilience" and "almost lack of resilience". (2) The evolution of the activity-environment system resilience is still absolutely volume driven, and the positive drive of relative quality is limited or even reversed. In addition, all the bilateral local spatial autocorrelations are generally represented as "center-periphery" separated spatial structures, but it is not always consistent with the overall trend. (3) For the governance of urban resilience, it is necessary to focus on common improvement and matching of absolute volume and relative quality in contents, orderly flow, accumulation and dispersion of supply and demand in ideas, and the strong support of new ideas and technologies in methods.

Study on the Characteristics of Urban Residents' Commuting Behavior and Influencing Factors from the Perspective of Resilience Theory: Theoretical Construction and Empirical Analysis from Nanjing, China.

In the transitional period of China's urbanization, commuting problems and demands are diversified and multi-level, so commuting research topics, viewpoints, and analysis paths should be organically combined to dynamically adapt to the complex commuting contradictions. Based on this, this paper introduces the resilience theory to improve the research paradigm of commuting behavior. Taking Nanjing, China as a case study, with the help of the survey data of commuting behavior of typical communities, this paper provides an empirical analysis of the characteristics and influencing factors of urban residents' commuting behavior from the perspective of resilience theory. The results show that: (1) in the face of commuting pressure, to a large extent, most commuters can still obtain commuting adaptability and a medium level or higher of commuting resilience; and (2) personal attributes, living and employment environment, and commuting environment all have significant heterogeneity effects on commuting pressure, commuting adaptability, and commuting resilience. From the perspective of resilience theory, the means of regulating commuting conflicts are flexible, which can not only directly reduce commuting pressure or optimize commuting adaptability, but also improve commuting resilience according to the specific commuting scenarios constructed by commuting pressure and adaptability. On the whole, the principles of comprehensive improvement, on-demand supply, and dynamic adjustment should be followed.

The Prevalence, Antibiotic Resistance and Biofilm Formation of Staphylococcus aureus in Bulk Ready-To-Eat Foods.

The prevalence of Staphylococcus aureus in 2160 bulk ready-to-eat foods from the Sichuan province of China during 2013-2016 was investigated. The antibiotic resistance and the associated genes, as well as biofilm formation capacity of the S. aureus isolates were measured. Furthermore, the relationship between the antibiotic resistance and the resistant genes was discussed. It was found that 54 S. aureus isolates were recovered, and their prevalence in meat products, dairy, fruit and vegetables, and desserts were 31 (2.6%), six (3.0%), nine (2.2%) and eight (2.3%), respectively. Most strains (52/54) were resistant to at least one of the antibiotics, and 21 isolates were identified as multidrug-resistant (MDR) S. aureus. Three isolates were found to be methicillin-resistant S. aureus. Penicillin, erythromycin, clindamycin, tetracycline and inducible clindamycin resistance were determined as the predominant antibiotics, and the isolates with the phenotypic resistance on these five antibiotics were all determined positive for the resistant gene associated. In total, 33 of 54 S. aureus isolates showed biofilm formation capacity, including two strong biofilm producers, one moderate and 30 weak ones. Two S. aureus isolates with strong biofilm formation abilities showed multi-drug resistance, and one moderate biofilm producer was resistant to two categories of antibiotics.

Interaction between rpsL and gyrA mutations affects the fitness and dual resistance of Mycobacterium tuberculosis clinical isolates against streptomycin and fluoroquinolones.

BACKGROUND: The interaction between different drug-resistant mutations is important to the development of drug resistance and its evolution. In this study, we aimed to reveal the potential relationships between mutations conferring resistance to two important antituberculosis drugs streptomycin (STR) and fluoroquinolones (FLQ). MATERIALS AND METHODS: We used an in vitro competitive fitness assay to reveal the interactions between different mutations of rpsL and gyrA in drug-resistant Mycobacterium smegmatis, followed by the analysis of the frequency of rpsL and gyrA mutation combinations in 213 STR-FLQ dual-resistant clinical Mycobacterium tuberculosis isolates from Sichuan region, which was also investigated by the whole genome data from 3,056 global clinical M. tuberculosis isolates. RESULTS: The strains with K43R and K88R mutation in rpsL showed no difference in relative fitness compared with their susceptible ancestor, while K43N, K43M, K43T, and K88E exhibited a significantly lower relative fitness (P<0.05). For the FLQ-resistant mutants, all mutation types showed no difference in their relative fitness. Among STR-FLQ dual-resistant M. smegmatis strains, a lower fitness was detected in those with K43N/M/T and K88E instead of K43R and K88R mutations in rpsL. Among M. tuberculosis isolates harboring rpsL and gyrA dual mutations, the most two frequent combinatorial mutation types were K43R/D94G (n=37) and K43R/A90V (n=24), with the former being the most frequent one by both in vitro tests and clinical survey. CONCLUSION: Our results suggest that the interaction between rpsL and gyrA mutations affects the fitness cost in STR-FLQ dual-resistant M. smegmatis and also the predilection of mutation combinations in clinical M. tuberculosis isolates.

Chinese ethnic meat products: Continuity and development.

With their distinctive sensory characterizations and unique processing technologies, Chinese ethnic meat products possess great potential for development and continuity in modern China's meat industry. Due to the greater demand for meat products and higher quality and safety concerns in economically fast growing China, the development and continuity of ethnic meat products face its own unique challenges. In this review, the classification of typical ethnic products and their characteristics, and the research progress on their quality and processing technologies are discussed. The application of innovative and green technologies to improve the safety and quality of ethnic meat products for greater industrialization and sustainable development is highlighted. Furthermore, the strategy for promoting the production of Chinese ethnic meat products during the next five years is presented.

Characterization of mutations in streptomycin-resistant Mycobacterium tuberculosis isolates in Sichuan, China and the association between Beijing-lineage and dual-mutation in gidB.

Mutations in rpsL, rrs, and gidB are well linked to streptomycin (STR) resistance, some of which are suggested to be potentially associated with Mycobacterium tuberculosis genotypic lineages in certain geographic regions. In this study, we aimed to investigate the mutation characteristics of streptomycin resistance and the relationship between the polymorphism of drug-resistant genes and the lineage of M. tuberculosis isolates in Sichuan, China. A total of 227 M. tuberculosis clinical isolates, including 180 STR-resistant and 47 pan-susceptible isolates, were analyzed for presence of mutations in the rpsL, rrs and gidB loci. Mutation K43R in rpsL was strongly associated with high-level streptomycin resistance (P < 0.01), while mutations in rrs and gidB potentially contributed to low-level resistance (P < 0.05). No general association was exhibited between STR resistance and Beijing genotype, however, in STR-resistant strains, Beijing genotype was significantly correlated with high-level STR resistance, as well as the rpsL mutation K43R (P < 0.01), indicating that Beijing genotype has an evolutionary advantage under streptomycin pressure. Notably, in all isolates of Beijing genotype, a dual mutation E92D (a276c) and A205A (a615g) in gidB was detected, suggesting a highly significant association between this dual mutation and Beijing genotype.

Suitability of IS6110-RFLP and MIRU-VNTR for differentiating spoligotyped drug-resistant mycobacterium tuberculosis clinical isolates from Sichuan in China.

Genotypes of Mycobacterium tuberculosis complex (MTBC) vary with the geographic origin of the patients and can affect tuberculosis (TB) transmission. This study was aimed to further differentiate spoligotype-defined clusters of drug-resistant MTBC clinical isolates split in Beijing (n = 190) versus non-Beijing isolates (n = 84) from Sichuan region, the second high-burden province in China, by IS6110-restriction fragment length polymorphism (RFLP) and 24-locus MIRU-VNTRs. Among 274 spoligotyped isolates, the clustering ratio of Beijing family was 5.3% by 24-locus MIRU-VNTRs versus 2.1% by IS6110-RFLP, while none of the non-Beijing isolates were clustered by 24-locus MIRU-VNTRs versus 9.5% by IS6110-RFLP. Hence, neither the 24-locus MIRU-VNTR was sufficient enough to fully discriminate the Beijing family, nor the IS6110-RFLP for the non-Beijing isolates. A region adjusted scheme combining 12 highly discriminatory VNTR loci with IS6110-RFLP was a better alternative for typing Beijing strains in Sichuan than 24-locus MIRU-VNTRs alone. IS6110-RFLP was for the first time introduced to systematically genotype MTBC in Sichuan and we conclude that the region-adjusted scheme of 12 highly discriminative VNTRs might be a suitable alternative to 24-locus MIRU-VNTR scheme for non-Beijing strains, while the clusters of the Beijing isolates should be further subtyped using IS6110-RFLP for optimal discrimination.

Characterization of rifampin-resistant isolates of Mycobacterium tuberculosis from Sichuan in China.

Rifampin is a key component of standard short-course first-line therapy against Mycobacterium tuberculosis, and rifampin resistance of this pathogen has been reported to be related to rpoB gene mutations. The objective of this study was to characterize the rpoB gene mutations in rifampin-resistant M. tuberculosis isolates circulated in Sichuan. Sequencing of rpoB gene and spoligotyping were performed on 268 randomly selected isolates from January 2008 to May 2010. The results indicated that 207 (97.2%) rifampin-resistant isolates had mutations at 26 codons in the amplified region with 50 different genotypes, while 1 (1.8%) of 55 susceptible isolates had a nonsense mutation. The most common mutations were in codon 531 (55.9%), 526 (16.4%), 516 (10.3%) and 511 (8.9%). Among the 213 resistant isolates, 150 (70.4%) belonged to the Beijing family and mutation at codon 531 (TCG→TTG) was associated with Beijing genotype (χ(2), 9.8305; p, 0.0017). It is demonstrated that the frequency of 511 (CTG→CCG) mutations in Sichuan was higher than in other provinces of China, as well as other geographic regions worldwide. It is suggested that only a small portion (2.7%) of rifampin-resistant Beijing genotype isolates in Sichuan be spread by clonal expansion during the study period.

The population structure of drug-resistant Mycobacterium tuberculosis clinical isolates from Sichuan in China.

China ranks second next to India among 22 high-burden countries despite decades' effort on tuberculosis (TB) control. The Sichuan province today contains the second-largest number of TB cases among Chinese provinces, where the prevalence of drug-resistant TB, especially MDR-TB, is much higher than the average level in eastern China. In this study, the population structure and the transmission characteristics of drug-resistant TB in Sichuan province were studied by spoligotyping and 24-locus Mycobacterial interspersed repetitive units-variable number tandem DNA repeats (MIRU-VNTR), applied to a total of 306 clinical isolates. Spoligotyping-based analysis showed that Beijing family represented 69.28% of all isolates and constituted the largest group (66.24%) of MDR-TB in Sichuan. The remaining isolates, accounting for 33.76% of MDR isolates, belonged to the ill-defined T family, Manu2, H3, LAM9, and other minor unassigned clades. The discriminatory power evaluated for spoligotyping was poor (HGI=0.595), but high for 24-locus MIRU-VNTRs (HGI=0.999). The number of the most discriminatory loci (h>0.6) was 12, including locus 424, 802, 960, 1644, 1955, 2163b, 2996, 3007, 3192, 3690, 4348 and 4052. It was concluded that 24-locus MIRU-VNTRs could be a more discriminatory tool for differentiating clinical isolates from Sichuan region. The small clustering size obtained from the current population structure analysis suggested that the high prevalence of drug-resistant TB in this region might be attributed partially to the acquired resistance due to inappropriate drug use rather than active transmission of drug-resistant TB (primary resistance).