[Relationship Between the VNTR Loci Polymorphism and MALDI-TOF MS Protein Profile Diversity in Mycobacterium tuberculosis Complex].

OBJECTIVE: To study the relationship between the copy numbers of repetitive units at variable number tandem repeat (VNTR) loci of Mycobacterium tuberculosis complex (MTBC) with its diversity of protein profiles. METHODS: The MTBC strains were subjected to genotyping using multiple locus variable number tandem repeat analysis (MLVA). Also, the principal component analysis (PCA) was performed for bacterial protein profiles of MTBC using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The relationship between the polymorphism of VNTR loci and PCA clustering was analyzed. RESULTS: A total of 157 MTBC strains were collected. 146 MTBC strains (MS identification score values ≥1.700) were performed PCA and three clusters, clusterⅠ(61 strains), clusterⅡ(26 strains) and cluster Ⅲ(59 strains), were generated. Polymorphic diversities were observed in 24 VNTR loci, among them, 7 were highly various, 7 were moderately, and 10 were low various. The polymorphism of Mtub39, QUB26 and QUB4156 loci were correlated with the results of MALDI-TOF MS clustering (P=0.000, P=0.035, P=0.017). CONCLUSION: The polymorphism of Mtub39, QUB26 and QUB4156 loci in MTBC was correlated with the difference of MALDI-TOF MS protein profiles, suggesting that these loci may play a role in regulating the composition of protein profiles of MTBC strains.

[Triton X-100 Enhanced the Detection of Hodge Test and Carba NP Test for Carbapenemase-producing Acinetobacter baumannii Complex].

OBJECTIVE: To find out highly effective phenotypic methods to detect carbapenemase-producing Acinetobacter baumannii (A. baumannii complex) so as to support the epidemiological investigation and clinical application. METHODS: We included 113 A. baumannii complex and compared the detection performance of modified Hodge test, Carba NP test, Triton Hodge test, and the simplified Carba NP-direct test with Tritont X-100. RESULTS: We tested 83 carbapenemase-producing A. baumannii complex and 30 non-carbapenemase producers. The sensitivity and specificity of Hodge test were significantly higher than those of Carba NP test (71.1% versus 35.0%, 100% versus 86.7%, P<0.05, respectively). The sensitivity of Triton Hodge test and Carba NP-direct test was respectively significantly higher than Hodge test and Carba NP test (98.8% versus 71.1%, 85.5% versus 35.0%, P<0.001, respectively ). However, the specificities were comparable (P>0.05). The positive additive effects of the two methods with Triton X-100 were more obvious than those of the methods without Triton X-100 (P<0.001). CONCLUSION: Triton X-100 could increase the sensitivity and positive additive effect on phenotypic detection of A. baumannii complex. Triton Hodge test and Carba NP-direct test were more applicable for clinical routine procedure.

Rapid antimicrobial susceptibility testing by matrix-assisted laser desorption ionization-time of flight mass spectrometry using a qualitative method in Acinetobacter baumannii complex.

The transmission and infections of multidrug-resistant bacteria can be prevented by rapid identification and antibiotic susceptibility testing (AST) for pathogenic bacteria in a clinical microbiology laboratory. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been routinely used as a tool for the identification of pathogens; however, a simple and accurate method for a rapid determination of the antimicrobial susceptibility profile is an urgent requisite. The present study established a method based on mass spectrometry to determine the drug resistance. Acinetobacter baumannii complex isolates were tested as an example. After short-term culture, the isolates were incubated with meropenem of different concentrations to determine the growth or the inhibition of the growth by MALDI-TOF MS. The agreement of minimum inhibitory concentration (MIC) values between MALDI-TOF MS-based rapid AST and broth microdilution method in susceptible and resistant strains was 77.1% and 70.1%, respectively. The susceptibility-breakpoint concentration (2 µg/mL) achieved a 98.9% sensitivity and 100% specificity with respect to resistance detection. Similarly, 96.9% sensitivity and 100% specificity were obtained for resistance detection with meropenem concentration at 8 µg/mL. MALDI-TOF MS-based rapid AST was applied to determine the drug resistance at breakpoint concentration, although MS-MICs might shift to a low dilution. Thus, it is critical for patients to accelerate the AST result from two days to several hours.

Extended-spectrum ß-lactamase-producing E. coli septicemia among rectal carriers in the ICU.

The aim of this study was to identify risk factors for extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli (E coli) bloodstream infection (BSI) among carriers hospitalized between March 2011 and June 2016 at the ICU of the West China Hospital.The cases were patients with at least 1 episode of ESBL-producing E coli BSI within 1 week after a positive rectal swab. Controls were selected randomly 1:2 among ESBL-producing E coli rectal carriers who did not develop BSI.Among 19,429 ICU patients, 9015 (46.4%) had a positive rectal swab for ESBL-producing E coli. Of them, 42 (0.5%) were diagnosed with ESBL-producing E coli BSI. The in-hospital mortality was higher for the BSI patients compared with controls (19.1% vs. 6.0%, P = .031). In the past 72 hours, patients in case group were more likely to use penicillin (odds ratio [OR] = 12.076; 95% confidence interval [CI]: 1.397-104.251, P = .02), cephalosporin (OR = 6.900; 95% CI: 1.493-31.852, P = .01), and carbapenem (OR = 5.422; 95% CI: 1.228-23.907, P = .03) as compared to patients in control group. Also, when compared to patients in control group, patients in case group were likely to stay for a longer time in ICU before positive rectal swab test (OR = 1.041, 95% CI: 1.009-1.075, P = .01) and have higher maximum body temperature before positive rectal swab (OR = 8.014; 95% CI: 2.408-26.620, P = .001).Bacteremia owing to ESBL-producing E coli was associated with high antimicrobial exposure, hospital stay, and maximum body temperature.