Efficacy of a membrane concentration method combined with real-time PCR for detection of Giardia and Cryptosporidium in drinking water.

The occurrence of Giardia and Cryptosporidium (oo)cysts in drinking source water poses a serious public health risk. Here, we established a method that combines membrane concentration and real-time polymerase chain reaction (PCR) to quantify Giardia and Cryptosporidium in drinking water. The water samples were filtered through a cellulose membrane to collect Giardia and Cryptosporidium, and then nucleic acids were extracted. Specific primers and probes were designed and synthesized according to the gph gene sequence of Giardia and 18S rRNA gene sequence of Cryptosporidium. The concentrations of the two targets were determined using real-time PCR technology. The sensitivity, specificity, and stability of the method were evaluated. Our findings revealed that the detection limits of real-time PCR method for detecting Giardia and Cryptosporidium were 0.926 and 0.65 copy/µL, respectively; the spiked recovery rates were above 60% and 38%, respectively, and relative standard deviations were under 0.95% and 2.26%, respectively. Therefore, this effective procedure based on the membrane concentration method and real-time PCR will be useful for detecting Giardia and Cryptosporidium in drinking water for purpose of continuous environmental monitoring.

Pressure response of carbapenems Klebsiella pneumoniae under antibiotic stress.

To analyze the drug-resistant phenotype and genetic characteristics of Carbapenem resistant Klebsiella pneumoniae (CRKP) in this region, and to study its different expression profiles in RNA level under the pressure of low levels of antibiotics. Trace dilution method and PCR method were used to detect the antibiotic resistance phenotype and antibiotic resistance gene carrying of CRKP strain, simulate the antibiotic stress process, and RNAseq was used to analyze the transcriptomic changes of CRKP strain. 37 CRKP strains, 27 Carbapenem sensitive Klebsiella pneumoniae (CSKP) CSKP strains and 42 sensitive strains were detected. The antibiotic resistance rate of CRKP strain was significantly higher than that of other drug-resistant strains, and there were many kinds of antibiotic resistance genes. Transcriptomic analysis showed that CRKP strain showed compensatory rise under meropenem stress at low concentration, and the expression of genes related to biofilm formation, pressure induction, pressure tolerance and transcriptional regulation was significantly changed. It was speculated that mrkAB, fimDH, phoHP and pspABCD clusters significantly altered their expression under the antibiotics stress response in CRKP strain. The detection rate of CRKP strain is high in this area. Under low levels of antibiotic stress, CRKP strain can not only survive by synthesizing antibiotic modified enzyme, but also respond by transcriptional regulation and biofilm changes, resulting in stress compensation. The discovery of this phenomenon explains the failure of treatment due to improper use of higher-order antibiotics from the perspective of genetic interaction.

Application advantages of liquid phase suspension chip technique in Salmonella Serotyping / 公共卫生与预防医学

Objective To probe into the feasibility and applicability of the foodborne food risk monitoring and sentinel hospital foodborne disease surveillance on the Salmonella serotype identification by liquid phase suspension array. Methods The serotyping of Salmonella isolates was performed by traditional glass agglutination test. Meanwhile, the liquid phase suspension array was operated to analyze the antigen O, H and AT for classification identification. Results From 2012 to 2019, a total of 215 strains of Salmonella were collected divided into 38 serotypes, 96% of them could be analyzed by SSA kit. The results of xMAP were in accordance with the traditional agglutination. 8 of the 11 strains which cannot be checked out by glass agglutination seemed to be easy detected by liquid phase suspension array. Conclusion The liquid phase suspension array has advantages of high throughput, high sensitivity and high specificity. It is able to detect the Salmonella serotype rapidly during a short time. Compared with the traditional serum agglutination method, the liquid phase suspension array has obvious advantages in detection time. It can be useful and important in the break out of foodborne disease caused by Salmonella spp.

Analysis of serum and drug resistance levels between food source and human salmonella / 公共卫生与预防医学

Objective To analyze serotype distribution, drug resistance, quinolone resistance gene carrying status and genetic relationship of foodborne Salmonella and human Salmonella isolates in Changzhou from 2012 to 2018, to provide scientific basis for the prevention and control of Salmonella. Methods The serum type was identified by serum agglutination and liquid chip. The antibiotic sensitivity was determined by micro broth dilution method. The quinolone antibiotic resistance gene was determined by gene sequencing method. The multilocus sequence typing ( MLST ) typing was performed on quinolone-resistant Salmonella, and the genetic relationship was analyzed by BioNumerics 8.0. Results A total of 10 and 36 serotypes were detected in 46 foodborne Salmonella strains and 152 human Salmonella strains, respectively. The dominant serotypes were Indiana Salmonella and Salmonella typhimurium. Erythromycin resistance rate was the highest in both Salmonella strains, and the proportion of multidrug-resistant bacteria was 93.47 % ( 43 / 46 ) and 80.92 % ( 123 / 152 ), respectively. 38 strains of quinolone-resistant foodborne Salmonella GyrA subunit mainly occurred double mutations Asp87Asn, Ser83Phe, ParC subunit mainly occurred single mutation Ser80Arg, 119 strains of quinolone-resistant human Salmonella qnrS gene detection rate was higher, reached 68.1 % ( 81 / 119 ) ; The dominant ST types of quinolone-resistant Salmonella from two sources were ST17 and ST19, respectively. Conclusions The antibiotic sensitivity of the two Salmonella resistant strains from Changzhou was the same ; Synergistic drug resistance, but both quinolone resistance genemutations and carry inconsistent ; The ST type distribution of quinolone resistant strains isalso inconsistent, and the genetic relationship is far. It is suggested that the probability of Salmonella resistant bacteria infection caused by food transmission in our region is small, and the treatment of the two should be differentiated.

The application of MALDI-TOF mass spectrometry in pathogen detection from the nosocomial infection / 重庆医学

Objective Pathogens from the nosocomial infection have been analyzed by MALDI‐TOF microbial identification system ,to evaluate mass spectrometry analysis advantage and explore the mass spectrometry method .Methods The pathogens have been analyzed by MALDI‐TOF microbial identification system ,by compared with the VITEK‐2 compact detection in the tes‐ting time ,detection rate and the amounts of identified strains .The homology differences have been analyzed by comparison calcula‐tion of common peaks from the fingerprint spectrums .Results Thirty‐one Escherichia coli strains ,28 Klebsiella pneumonia strains and 9 unusual pathogen strains have been identified by MALDI‐TOF MS for only 1 hours .It has more advantages than VITEK‐2 in the testing time and other aspects .Conclusion Nosocomial infection of pathogen shows a point source propagation mode centering on the department .MALDI‐TOF mass spectrometry is able to rapidly and correctly identify the pathogen .MALDI‐TOF microbial i‐dentification system is expected to be the major detecting technique in the field of the pathogen monitor and resistance monitoring a ‐nalysis .