The Association of Serum hsCRP and Urinary Alpha1-Microglobulin in Patients with Type 2 Diabetes Mellitus.

This study aimed to investigate the association of serum hsCRP and urinary A1MG in patients with T2DM. Numerous investigations have proven that serum hypersensitive C-reactive protein (hsCRP) concentration in patients with type 2 diabetes mellitus (T2DM) is increased. Also, increased urinary alpha-1 microglobulin (A1MG) can be an early sign of renal damage, primarily on the proximal tubules in T2DM. Little information is available with respect to the associations of serum hsCRP levels and urinary A1MG in T2DM. A total of 520 patients with T2DM were recruited to participate in this study. Serum hsCRP and UA1MG (urinary alpha1-microglobulin to creatinine ratio), UACR (urinary microalbumin to creatinine ratio), UIGG (urinary immunoglobulin G to creatinine ratio), and UTRF (urinary transferrin to creatinine ratio) were obtained. The association of serum hsCRP level and each urinary protein parameter was analyzed by using the regression analysis, respectively. LnhsCRP was positively associated with the lnUA1MG in all three linear regression models (adjusted ß in model 3=0.122, SE=0.027, P<0.001). Furthermore, the high hsCRP group (hsCRP > 3mg/L) was associated with increasing risk of high UA1MG (adjusted OR in model 3=1.610, 95% CI 1.037-2.499, P=0.034) by logistic regression. This study suggests that serum hsCRP levels independently associate with UA1MG in patients with T2DM. Further research is warranted to elucidate these interactions.

Epidemiology and antifungal susceptibilities of yeast isolates causing invasive infections across urban Beijing, China.

AIM: To investigate the species distribution and antifungal susceptibility profiles of yeast isolates causing invasive infections across Beijing. MATERIALS & METHODS: A total of 1201 yeast isolates recovered from blood and other sterile body fluids were correctly identified by matrix-assisted laser desorption/ionization TOF MS supplemented by DNA sequencing. Antifungal susceptibility testing was performed according to the Clinical and Laboratory Standards Institute broth microdilution method. RESULTS: Candida (95.5%) remained the most common yeast species isolated; Candida albicans (38.8%) and Candida parapsilosis (22.6%) were the leading species of candidemia. Azole resistances were mainly observed in Candida glabrata and Candida tropicalis isolates. CONCLUSION: This study outlined the epidemiologic data of invasive yeast infections and highlighted the need for continuous monitoring of azole resistances among C. glabrata and C. tropicalis isolates in Beijing.

Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry systems for the identification of clinical filamentous fungi.

Infections caused by filamentous fungi have become a health concern, and require rapid and accurate identification in order for effective treatment of the pathogens. To compare the performance of two MALDI-TOF MS systems (Bruker Microflex LT and Xiamen Microtyper) in the identification of filamentous fungal species. A total of 374 clinical filamentous fungal isolates sequentially collected in the Clinical Laboratory at the Beijing Tongren Hospital between January 2014 and December 2015 were identified by traditional phenotypic methods, Bruker Microflex LT and Xiamen Microtyper MALDI-TOF MS, respectively. The discrepancy between these methods was resolved by sequencing for definitive identification. Bruker Microflex LT and Xiamen Microtyper had similar correct species ID (98.9 vs. 99.2%), genus ID (99.7 vs. 100%), mis-ID (0.3 vs. 0%) and no ID (0 vs. 0). The rate of correct species identification by both MALDI-TOF MS (98.9 and 99.2%, respectively) was much higher compared with phenotypic approach (91.9%). Both MALDI-TOF MS systems provide accurate identification of clinical filamentous fungi compared with conventional phenotypic method, and have the potential to replace identification for routine identification of these fungi in clinical mycology laboratories. Both systems have similar performance in the identification of clinical filamentous fungi.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid identification of fungal rhinosinusitis pathogens.

PURPOSE: Filamentous fungi are among the most important pathogens, causing fungal rhinosinusitis (FRS). Current laboratory diagnosis of FRS pathogens mainly relies on phenotypic identification by culture and microscopic examination, which is time consuming and expertise dependent. Although matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS has been employed to identify various fungi, its efficacy in the identification of FRS fungi is less clear. METHODOLOGY: A total of 153 FRS isolates obtained from patients were analysed at the Clinical Laboratory at the Beijing Tongren Hospital affiliated to the Capital Medical University, between January 2014 and December 2015. They were identified by traditional phenotypic methods and Bruker MALDI-TOF MS (Bruker, Biotyper version 3.1), respectively. Discrepancies between the two methods were further validated by sequencing. RESULTS: Among the 153 isolates, 151 had correct species identification using MALDI-TOF MS (Bruker, Biot 3.1, score ≥2.0 or 2.3). MALDI-TOF MS enabled identification of some very closely related species that were indistinguishable by conventional phenotypic methods, including 1/10 Aspergillus versicolor, 3/20 Aspergillus flavus, 2/30 Aspergillus fumigatus and 1/20 Aspergillus terreus, which were misidentified by conventional phenotypic methods as Aspergillus nidulans, Aspergillus oryzae, Aspergillus japonicus and Aspergillus nidulans, respectively. In addition, 2/2 Rhizopus oryzae and 1/1 Rhizopus stolonifer that were identified only to the genus level by the phenotypic method were correctly identified by MALDI-TOF MS. CONCLUSION: MALDI-TOF MS is a rapid and accurate technique, and could replace the conventional phenotypic method for routine identification of FRS fungi in clinical microbiology laboratories.

Rapid low-cost detection of Klebsiella pneumoniae carbapenemase genes by internally controlled real-time PCR.

An internally controlled real-time PCR assay based on SYBR Green was developed to screen Klebsiella pneumoniae carbapenemases (KPCs) gene containing bacteria and was validated for clinical strains or surveillance specimens. When 248 clinical samples were tested, the sensitivity and specificity of the assay were 100% and 99%, respectively.

[Risk factors for acquired multidrug-resistant Acinetobacter baumannii colonization in respiratory intensive care unit].

OBJECTIVE: To determine the risk factors for respiratory intensive care unit (RICU)-acquired colonization of multidrug-resistant Acinetobacter baumannii (MDR-AB). METHODS: From January 2010 to June 2011, active screening was performed to define patients with RICU-acquired colonization of MDR-AB. And environment surveillance was carried out and patient data were collected. Logistic regression was applied to identify the risk factors of RICU-acquired colonization of MDR-AB. RESULTS: Active screening for MDR-AB was performed for 110 patients in RICU and 50 patients turned out to be positive. After eliminating 3 input positive patients, the RICU-acquired colonization rate of MDR-AB was 43.9% (47/107). The environmental contaminated rate of MDR-AB was 66.0% (31/47) for 47 positive patients and 33.9% (19/56) for 56 negative ones (χ(2) = 10.494, P < 0.01). Five risk factors were associated with the colonization of MDR-AB through univariate analysis: consciousness disturbance, use of carbapenems, nasal feeding tube, endotracheal intubation and mechanical ventilation (all P < 0.05). The Logistic regression equation contained 3 risk factors of conscious disturbance, use of carbapenems and mechanical ventilation (OR = 3.412, 3.211, 3.002; 95% CI: 1.165 - 9.992, 1.117 - 9.233, 1.101 - 8.182). CONCLUSION: Three risk factors are independently associated with the RICU-acquired colonization of MDR-AB: consciousness disturbance, use of carbapenems and mechanical ventilation.

A rapid low-cost real-time PCR for the detection of Klebsiella pneumonia carbapenemase genes.

BACKGROUND: Klebsiella pneumonia carbapenemases (KPCs) are able to hydrolyze the carbapenems, which cause many bacteria resistance to multiple classes of antibiotics, so the rapid dissemination of KPCs is worrisome. Laboratory identification of KPCs-harboring clinical isolates would be a key to limit the spread of the bacteria. This study would evaluate a rapid low-cost real-time PCR assay to detect KPCs. METHODS: Real-time PCR assay based on SYBR GreenIwas designed to amplify a 106 bp product of the blaKPC gene from the 159 clinical Gram-negative isolates resistant to several classes of -lactam antibiotics through antimicrobial susceptibility testing. We confirmed the results of real-time PCR assay by the conventional PCR-sequencing. At the same time, KPCs of these clinical isolates were detected by the modified Hodge test (MHT). Then we compared the results of real-time PCR assay with those of MHT from the sensitivity and specificity. Moreover, we evaluated the sensitivity of the real-time PCR assay. RESULTS: The sensitivity and specificity of the results of the real-time PCR assay compared with those of MHT was 29/29(100%) and 130/130(100%), respectively. The results of the real-time PCR and the MHT were strongly consistent (Exact Sig. (2-tailed) =1. 000; McNemar test). The real-time PCR detection limit was about 0.8 cfu using clinical isolates. CONCLUSION: The real-time PCR assay could rapidly and accurately detect KPCs -harboring strains with high analytical sensitivity and specificity.

[Performance of locked nucleic acid probe real-time polymerase chain reaction in the detection of Aspergillus fumigatus].

OBJECTIVE: To evaluate the performance of locked nucleic acid (LNA) probe Real-time polymerase chain reaction (PCR) in the detection of Aspergillus fumigatus (A. fumigatus). METHODS: All clinically cultured isolates of Aspergillus at our hospital were identified by morphology and DNA sequencing assay. The experimental group consists of A. fumigatus (n = 48) while the control group was made up of A. flavus (n = 55), A. versicolor (n = 16), A. nidulans (n = 10), A. sydowii (n = 5) and A. parasiticus (n = 1). The clinical samples consisted of A. fumigatus sinusitis tissue (n = 20) and bronchoalveolar lavage fluid (n = 1). DNA was extracted from all samples. A. fumigatus ß-tubulin gene was targeted with LNA probe Real-time PCR assay. LNA probe Real-time PCR was evaluated with regards to specificity, efficiency, linear dynamic range in PCR amplification and limits of detection. RESULTS: All clinical samples were positively amplified. The specificity was 100% and the PCR efficiency 98.2%. Linear dynamic range was at least six orders of magnitude and the limit of detection 2.5 pg. CONCLUSION: LNA probe Real-time PCR is a promisingly accurate assay of rapidly detecting A. fumigatus practically and cost-efficiently.

[Correlation factor analysis of pan-drug resistant Acinetobacter baumannii strains in acquired infections].

OBJECTIVE: To explore the clinical factors, drug resistance and molecular epidemiology homologous characteristics of pan-drug resistant Acinetobacter baumannii (PDRAB) in acquired infections and analyze the correlation factor between epidemic characteristics and acquired infections. METHODS: A total of 60 PDRAB strains from nine acquired infections and related clinic data were collected from January 2009 to January 2011. The drug-resistant phenotype was tested by disk diffusion methods. The isolate identification and homology were studied by automation repetitive-element sequence-based (REP)-PCR typing platform from genes and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF MS) from proteins. RESULTS: All strains were resistant to 12 antibiotics except 2 strains to imipenem and meropenem. The strains in this study were divided into 12 types (A-L) by REP-PCR. And 60 strains were also clustered to a-e types by MALDI-TOF-TOF MS. Compared with MALDI-TOF-TOF MS, REP-PCR tended to be more accurate. Breathing machine carriage and cross transmission were the main reasons for a major epidemic outbreak at department of pulmonary medicine from July 2009 to October 2009. Hand transmission of medical care personnel was a key factor for SICU 2010 January to February. The contamination and transmission to environment of PDRAB in nasal pharynx or respiratory tract by superspreader were the main reasons for the other 7 epidemic outbreaks. Department of emergency medicine was the source of acquired infections. CONCLUSION: The key control measures of acquired infections are early identification and isolation of spreader, environment and instrument disinfection, hand washing and rational uses of antibiotics. MALDI-TOF-TOF MS will become a preferred tool of identification and classification of microorganisms because of its simple operation, affordable price and handling rapidity.