Genomic characterization of Campylobacter isolates in Huzhou, China.

Campylobacter is a major foodborne pathogen that causes outbreaks and sporadic gastrointestinal disease, creating a serious disease burden. Campylobacter strains isolated from diarrhea cases (n = 11) and raw poultry meat products (n = 2) in Huzhou, including 11 Campylobacter jejuni and 2 Campylobacter coli strains, were subjected to virulence gene, drug resistance gene, genetic correlation, antibiotic resistance, and multilocus sequence typing (MLST) analyses. The 13 Campylobacter isolates were divided into 12 sequence types (STs), one of which was a new ST. The isolated strains contain multiple virulence-related genes. Drug sensitivity analysis showed that the resistance rate of the 13 isolates to nalidixic acid, ciprofloxacin, and tetracycline was 92.3%. Genome sequencing indicated that all 11 strains of C. jejuni carried the tet(O) and blaOXA resistance genes, and 2 strains of C. coli carried multiple drug resistance genes. Phylogenetic analysis based on core-genome single-nucleotide polymorphisms indicated that the 11 C. jejuni isolates from diarrhea patients and food sources are not closely phylogenetically related.

Diagnostic accuracy of electronic surveillance tool for catheter-associated urinary tract infections in tertiary care hospitals: A meta-analysis.

BACKGROUND: Automated systems have been developed to reduce labor-intensive manual recordings during nosocomial infection surveillance. The diagnostic accuracies of these systems have differed in various settings. METHODS: We designed this meta-analysis to evaluate the diagnostic accuracy of an electronic surveillance tool for catheter-associated urinary tract infections (CAUTIs) in tertiary care hospitals. We systematically searched databases such as Medline, Scopus, Cochrane library and Embase (from inception until November 2019) for relevant studies. We assessed the quality of trials using the diagnostic accuracy studies-2 tool, and performed a meta-analysis to obtain a pooled sensitivity and specificity for electronic surveillance. We included 6 studies with 16,492 patients in the analysis. RESULTS: We found a pooled sensitivity of electronic diagnostic surveillance for CAUTIs of 97.5% (95% confidence interval [CI], 67.6-99.9%) and a pooled specificity of 92.6% (95% CI, 55.2-99.2%). The diagnostic odds ratio was 494 (95% CI, 89-2747). The positive likelihood ratio was 13.1 (95% CI, 1.63-105.8) and the negative likelihood ratio 0.02 (95% CI, 0.001-0.40). A bivariate box plot indicated the possibility of heterogeneity between the included studies. CONCLUSION: Our review suggests that electronic surveillance is useful for diagnosing CAUTIs among hospitalized patients in tertiary care hospitals due to its high sensitivity and specificity.

Emergence and predominance of norovirus GII.17 in Huzhou, China, 2014-2015.

BACKGROUND: Norovirus (NoV) has been recognized as the leading cause of both outbreaks and sporadic cases of acute gastroenteritis in children and adults worldwide. Stool samples collected from outpatients with clinical symptoms of acute gastroenteritis in all age groups at the First People's Hospital in Huzhou, Huzhou, China between March 2014 and February 2015 were analyzed to gain insight into the epidemiology and genetic variation in NoV strains circulating in China. METHOD: Real-time RT-PCR (qPCR) was performed for Norovirus detection. RT-PCR were used for genomic amplification and sequencing. Genogroup and genotype were assigned using the NoV Noronet typing tool and the strains were named according to the time of isolation. The phylogenetic analysis was conducted using MEGA 5. RESULTS: Of the 809 specimens, 193 (23.9 %) were positive for NoV, with GII.4 and GII.17 the most commonly identified strains. Phylogenetic analysis confirmed the presence of five recombinant strains in Huzhou. Recombinants GII.P13/GII.17 and GII.P12/GII.4 were newly detected in China. The GII.P13/GII.17 recombinant was first identified in October 2014 and steadily replaced GII.Pe/GII.4 (GII.4 Sydney 2012) as the predominant circulating NoV genotype. CONCLUSION: This is the first report of the detection of GII.17 in the Huzhou area and of a NoV genotype being detected in greater numbers than GII.4. Furthermore, our results indicated that following the emergence of GII.17 in October 2014, it steadily replaced the previous circulating GII.4 Sydney2012 strain, which was the dominant circulating genotype for the past 2 years. As norovirus are the important cause of nonbacterial gastroenteritis, continuous and comprehensive study of the norovirus strains involved in large and cost-effective acute gastroenteritis would help understanding the molecular epidemiology of norovirus infections and development of improved prevention and control measures.

A cross-priming amplification assay coupled with vertical flow visualization for detection of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a marine seafood-borne pathogen that causes gastrointestinal disorders in humans. In this study, we developed a cross-priming amplification (CPA) assay coupled with vertical flow (VF) visualization for rapid and sensitive detection of V. parahaemolyticus. This assay correctly detected all target strains (n = 13) and none of the non-target strains (n = 27). Small concentrations of V. parahaemolyticus (1.8 CFU/mL for pure cultures and 18 CFU/g for reconstituted samples) were detected within 1 h. CPA-VF can be applied at a large scale and can be used to detect V. parahaemolyticus strains rapidly in seafood and environmental samples, being especially useful in the field.

Prevalence and genetic diversity of noroviruses in adults with acute gastroenteritis in Huzhou, China, 2013-2014.

Norovirus (NoV) infection is the most common cause of nonbacterial acute gastroenteritis, which affects both adults and children. However, the molecular epidemiology of NoV in adults with acute gastroenteritis in China has not been investigated extensively. In this study, we investigated the occurrence of NoV infections and analyzed the genetic diversity of NoV in adults with acute gastroenteritis in Huzhou, China. A total of 796 fecal samples were collected from outpatients (≥16 years of age) between March 2013 and February 2014. Real-time RT-PCR was performed to detect NoV genogroups I (GI) and II (GII). For genotyping, the capsid and RNA-dependent RNA polymerase (RdRp) genes were partially amplified and sequenced for phylogenetic analysis. NoVs were detected in 26.51% (211/796) of the specimens, with GII being predominant, representing 96.20% of the NoV infections. At least nine genotypes were identified among GI and GII specimens, including GI.P2/GI.2, GI.P3/GI.3, GI.P4/GI.4, GII.Pe/GII.4 Sydney_2012, GII.P12/GII.3, GII.P7/GII.6, GII.P16/GII.13, GII.Pe, and GII.Pg (RdRp only). This is the first report of a GII.P16/GII.13 recombinant virus in adults in China. GII.Pe/GII.4 Sydney_2012 was the most prevalent genotype and the only GII.4 variant identified during the study period. Our findings suggested that NoV was a common causative agent of acute gastroenteritis in adults in Huzhou, China. During the study period, the NoVs circulating in adults in Huzhou were predominantly GII.4 Sydney_2012 variants and GII NoV recombinants.

Rapid emergence of novel GII.4 sub-lineages noroviruses associated with outbreaks in Huzhou, China, 2008-2012.

Infection caused by noroviruses (NoVs) is one of the most important causes of acute gastroenteritis in humans worldwide. To gain insight into the epidemiology of and genetic variation in NoV strains, stool samples collected from 18 outbreaks of acute gastroenteritis in Huzhou, China, between January 2008 and December 2012 were analyzed. Samples were tested for NoVs by real-time RT-PCR. Partial sequences of the RNA- dependent RNA polymerase (RdRp) and capsid gene of the positive samples were amplified by RT-PCR, and the PCR products were sequenced and used for phylogenetic analysis. NoVs were found to be responsible of 88.8% of all nonbacterial acute gastroenteritis outbreaks in Huzhou over the last 5 years. Genogroup II outbreaks largely predominated and represented 93% of all outbreaks. A variety of genotypes were found among genogroups I and II, including GI.4, GI.8, GII.4, and GII.b. Moreover, phylogenetic analyses identified two recombinant genotypes (polymerase/capsid): GI.2/GI.6 and GII.e/GII.4 2012 Sydney. GII.4 was predominant and involved in 8/10 typed outbreaks. During the study period, GII.4 NoV variants 2006b, New Orleans 2009, and Sydney 2012 were identified. This is the first report of the detection of GII.4 New Orleans 2009 variant, GII.e/GII.4 Sydney 2012 recombinant in outbreaks of acute gastroenteritis in China.

[Molecular characteristics of noroviruses causing outbreaks of acute gastroenteritis in Huzhou].

OBJECTIVE: To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou. METHODS: From 2008 to 2010, total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tested for Norovirus. Partial sequence of RNA dependent RNA polymerase (RdRp) of the positive samples were amplified by RT-PCR, then the PCR production were purified, sequenced and put into phylogenetic analysis. RESULTS: 50 of 119 specimens were positive for Norovirus by real-time RT-PCR. Out of those 50 Norovirus positive specimens, 9 were Norovirus Genogroup I (GI) positive, 35 were Norovirus Genogroup II (GII) positive, 6 was both Norovirus GI and GII positive. 12 PCR products for RdRp were selected for further studies on sequencing. Phylogenetic analysis revealed that the 5 GI norovirus isolates were belonged to genotype GI/2 and GI/3. Of the 7 GII norovirus isolates, 6 were belonged to genotype GII/4, 1 was belonged to genotype Glib. CONCLUSION: Norovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.

Analysis of 15 STR loci in Chinese population from Sichuan in West China.

Allele frequencies for 15 STR loci found in PowerPlex 16 System kit were determined in a sample of 200 unrelated individuals living in Sichuan area in west China. The values of observed heterozygosity (Ho), power of discrimination (PD), probability of paternity exclusion (PE) and polymorphism information content (PIC) were calculated. All loci were in accordance with Hardy-Weinberg equilibrium (p<0.05). The obtained frequency distributions were compared with other previously reported population data.

[A minisequencing technique for genotyping 12 Y-SNP and its genetic polymorphisms].

OBJECTIVE: The aim of the present study was to establish a rapid and robust assay used to simultaneously genotype SNPs by the single nucleotide primer extension (minisequencing) with the SNaPshot Kit and obtain the population genetic data in Chinese population in Sichuan. The analysis of single nucleotide polymorphisms (SNPs) is a promising application in forensic casework. METHODS: 12 Y-SNPs, which were SRY2627, SRY1532, M13, M20, SRY8299, Tat, M69, M9, 92R7, M17, M19 and M112, were multiple amplificated and the PCR products were pooled, Purified, and then used as templates for the minisequencing reaction with the commol/Lercially available SNaPshot Kit. Then the products of minisequencing reaction were detected by capillary elcetrophoresis. RESULTS: 78 genomic DNA individual samples from Sichuan, China and 5 semen stain samples from sexal criminal scene were analyzed and two haplotypes could be identified. CONCLUSION: A rapid method has been established to analyze the 12 Y-SNPs by multiplex PCR and minisequencing. It can be applied in forensic casework successfully.