Rapid screening for antigenic characterization of GII.17 norovirus strains with variations in capsid gene.

The emergence of the novel GII.17 Kawasaki 2014 norovirus variant raising the interest of the public, has replaced GII.4 as the predominant cause of noroviruses outbreaks in East Asia during 2014-2015. Antigenic variation of the capsid protein is considered as one of the key mechanisms of norovirus evolution. In this study, we screened a panel of GII.17 mutants. First, we produced norovirus P proteins using cell-free protein synthesis (CFPS) system, comparing the results to pure proteins expressed in a cell-based system. Next, we determined the binding capability of specific monoclonal antibody (mAb) 2D11 using a unique set of wild-type GII.17 strains. Results of the EIA involving a panel of mutant cell-free proteins indicated that Q298 was the key residue within loop 1. These data highlighted the essential residues in the linear antibody binding characteristics of novel GII.17. Furthermore, it supported the CFPS as a promising tool for rapidly screening mutants via the scalable expression of norovirus P proteins.

Prevalence of Chromosomally Located blaCTX-M-55 in Salmonella Typhimurium ST34 Isolates Recovered from a Tertiary Hospital in Guangzhou, China.

Nontyphoidal Salmonella (NTS) is one of the most prevalent bacterial causes of gastrointestinal infections worldwide. Meanwhile, the detection rate of CTX-M-55 ESBL-positive has increased gradually in China. To identify the molecular epidemiological and genomic characteristics of blaCTX-M-55-carrying nontyphoidal Salmonella (NTS) clinical isolates, a total of 105 NTS isolates were collected from a Chinese tertiary hospital. Antimicrobial susceptibility testing was performed to determine the resistance phenotype. Whole-genome sequencing and bioinformatics analysis were used to determine the antimicrobial resistance genes, serotypes, phylogenetic relationships, and the genetic environment of the blaCTX-M-55 gene. The results showed that among the 22 ceftriaxone resistant isolates, the blaCTX-M-55 was the most common ß-Lactamase gene carried by 14 isolates, including serotypes S. Typhimurium (10/14), S. Muenster (2/14), S. Rissen (1/14), and S. Saintpaul (1/14). Phylogenetic analysis shows that 10 blaCTX-M-55-positive S. Typhimurium ST34 isolates were divided into two clusters. The genetic relationship of isolates in each cluster was very close (≤10 cgMLST loci). The blaCTX-M-55 gene was located on the chromosome in 10 isolates, on IncI1 plasmid in three isolates, and IncHI2 plasmid in one isolate. In conclusion, the blaCTX-M-55 gene, mainly located on the chromosome of S. Typhimurium ST34 isolates, was the main driving force associated with the resistance of NTS to cephalosporins. Therefore, close attention to the clonal dissemination of blaCTX-M-55-carrying S. Typhimurium ST34 in clinical settings must be monitored carefully. IMPORTANCE ESCs are the first choice for treating NTS infections. However, ESBLs and AmpC ß-lactamases are the most typical cause for ESCs resistance. The CTX-M-55 ESBL-positive rate has gradually increased in the clinic in recent years. At present, the research about blaCTX-M-55-positive Salmonella mainly focuses on the foodborne animals or the environment while less on clinical patients. Thus, this study was carried out for identifying molecular epidemiological and genomic characteristics of blaCTX-M-55-carrying NTS clinical isolates. The results showed that the blaCTX-M-55 gene, mainly located on the chromosome of S. Typhimurium ST34 isolates from Conghua District, was the main driving force associated with the resistance of NTS to cephalosporins. Therefore, our work highlights the importance of monitoring the clonal dissemination of blaCTX-M-55-carrying S. Typhimurium ST34 in clinical settings.

The VP2 protein exhibits cross-interaction to the VP1 protein in norovirus GII.17.

Norovirus is a major cause of acute gastroenteritis worldwide. Like the major capsid protein (VP1), the minor capsid protein (VP2) also contains a hypervariable domain. Generally, a hypervariable domain is functionally driven. However, many functions of VP2 remain unknown and worth exploring. Without sufficient sequences and an available crystallographic model, it is difficult to explore VP2's mysteries. As a helper of stabilizing and coordinating the formation of virus-like particles (VLPs), we asked whether VP2 interacted with the major capsid protein (VP1) in GII.17 and if so, what the key interaction residues were. Here, we reported cross-interaction among four strains represented four clusters of GII.17, and the VP1 interaction domain of VP2 (174-179aa) was found. However, the VP1 interaction domain of VP2 was not universal in different clusters of GII.17. VP2 might evolve in a different pattern from VP1. Additionally, in contrast to previous reports, we found that VP2 localized in the cytoplasm. More possibilities of VP2 should be further explored.

Corrigendum: Molecular Epidemiology and Characteristics of CTX-M-55 Extended-Spectrum ß-Lactamase-Producing Escherichia coli From Guangzhou, China.

[This corrects the article DOI: 10.3389/fmicb.2021.730012.].

Association of fucosyltransferase 2 gene with norovirus infection: A systematic review and meta-analysis.

BACKGROUND: Norovirus is a leading cause of viral gastroenteritis outbreaks worldwide. Histo-blood group antigens (HBGAs) are important host attachment factors in susceptibility to norovirus. In this study, the association of FUT2 gene, which participates in the biosynthesis of HBGAs, with norovirus infection has been investigated. METHODS: All relevant studies on the associations of FUT2 gene with norovirus were retrieved from PubMed, Web of Science, Embase, and Cochrane Library databases. Odds ratios (ORs) and 95% confidence interval (CI) were used to analyze the extracted data. I2 statistic, sensitivity analysis and publication bias analysis were used to confirm the findings. Subgroup analyses were performed for races, genotypes, development degree of the countries, publication years, age and setting when heterogeneity was recorded. RESULTS: Twenty studies including 4066 participants were included for the meta-analysis. This analysis showed that there is a significant association between FUT2 gene and norovirus infection (OR = 3.02, 95%CI = 2.00-4.55, P < 0.001). Additionally, the ORs of norovirus infection among Chinese (OR = 4.49, 95%CI = 2.37-8.50, P < 0.001) were higher than those among Caucasian (OR = 3.23, 95%CI = 2.20-4.74, P < 0.001). CONCLUSIONS: The meta-analysis suggested that FUT2 gene is associated with susceptibility to norovirus infection.

Molecular Epidemiology and Characteristics of CTX-M-55 Extended-Spectrum ß-Lactamase-Producing Escherichia coli From Guangzhou, China.

In recent years, the CTX-M-55 extended-spectrum ß-lactamase (ESBL)-positive rate has gradually increased in the clinic. To identify the molecular epidemiology and characteristics of bla CTX-M -55-positive isolates, a total of 374 non-repetitive ESBL-producing Escherichia coli strains were collected from patients in two hospitals in Guangzhou, and 89 bla CTX-M -55-positive isolates were selected by CTX-M-1-group PCR amplification and confirmed by DNA sequencing. Whole-genome sequencing was used to analyze the resistance phenotype, plasmid types, phylogenetic relationships and genetic environment of the bla CTX-M -55 gene. Conjugation experiments and PCR were performed to confirm whether the plasmid harboring bla CTX-M-55 gene could be transferred. The results showed that all bla CTX-M-55-positive isolates were resistant to ceftriaxone, and 88.76 and 76.40% were resistant to ceftazidime and cefepime, respectively. The resistance rates to levofloxacin and sulfamethoxazole were 66.29 and 59.55%, respectively. However, the sensitivity rate of piperacillin/tazobactam, amoxicillin/clavulanate, and amikacin exceeded 90%. All bla CTX-M-55-positive isolates were sensitive to carbapenems. Thirty-two STs were detected in the bla CTX-M-55-positive isolates, among which the detection rate of ST1193 was relatively high (19.10%, 17/89), and other ST types were scattered. It remains to be seen whether ST1193 carrying the bla CTX-M -55 gene can become a popular clone strain in this region in the future. The plasmid types carrying the bla CTX-M -55 gene included IncI1, IncFII, IncFIC, IncFIB, IncHI2, IncI2, and IncX/Y, among which the IncI1 and IncFII plasmids were the main plasmids, accounting for 37.80 and 28.09%, respectively. Among them, 11 strains of the IncI1 plasmid existed in ST1193 strains. The bla CTX-M -55 gene was found on chromosomes of 13 isolates, and seemed to be increasing annually. Up to five distinct types of genetic environments surrounding the bla CTX-M -55 gene were analyzed. The most common structure was type II "ISEcp1-bla CTX-M -55-ORF477." In conclusion, whether ST1193, which carries bla CTX-M -55 gene, will be an epidemic clone of this region in the future remains to be concerned. The plasmids IncI1 and IncFII, and mobile elements such as ISEcp1 and IS26 may be the main factors leading to the spread and prevalence of CTX-M-55 genotypes.

Global prevalence of norovirus in cases of acute gastroenteritis from 1997 to 2021: An updated systematic review and meta-analysis.

BACKGROUND: The worldwide response towards the acute gastroenteritis epidemic was well known, but the absence of an updated systematic review of global norovirus epidemiology in cases of gastroenteritis existed. We aimed to conduct and update a systematic review and meta-analysis of studies assessing norovirus prevalence among gastroenteritis patients worldwide. METHODS: Four databases (PubMed, EMBASE, Cochrane Library, and Web of Science) were searched for epidemiological papers from 2014 to 2021 which applied the PCR method to access the prevalence of norovirus in acute gastroenteritis patients more than a full year. Statistical analysis was conducted using R-4.0.0 software. RESULTS: A total of 405 records with 842, 926 cases were included. The pooled prevalence of norovirus was 16% (95%CI 15, 17). Children under 5 years old were at a higher risk with norovirus. A higher prevalence was seen in South America (22%, 95% CI 18, 27), while other continents showed a similar result with the overall prevalence of norovirus. No association was found between national income level and norovirus prevalence. A gradient of decreasing prevalence was noticed from community (20%, 95% CI 16, 24) to outpatients (18%, 95% CI 16, 20) to hospital setting (included both in- and outpatients, 17%, 95% CI 16, 19) to inpatients (15%, 95% CI 13, 17). CONCLUSION: Norovirus were associated with 16% acute gastroenteritis globally. To fully understand the prevalence of norovirus worldwide, the continual surveillance of norovirus epidemics was required.

Chromosome and Plasmid Features of Two ST37 Clostridioides difficile Strains Isolated in China Reveal Distinct Multidrug Resistance and Virulence Determinants.

Clostridioides difficile ST37 is an emerging and prevalent multilocus sequence type and represents a lineage of clinical significance. This study aimed to characterize two epidemic C. difficile ST37 strains, CD161 and CDT4. CD161 acquires a chromosome and two distinct plasmids, pCD161-L, sharing high similarity with Clostridium phage, and pCD161-S, while CDT4 has a chromosome and a plasmid pCDT4 identical to pCD161-S. In the chromosome of both strains, three CdISt1-like elements and a skinCd element, which might influence sporulation, were identified. The multidrug resistance of the strains was due to the mutation in 23S rDNA, gyrA, and gyrB genes and the acquisition of ermB, ant6-Ia, aac6'-aph2'', and tetM genes. In addition, a distinct pathogenicity locus (PaLoc) with truncated tcdA gene represents the genetic feature of ST37 strains. To our knowledge, this is the first complete genome, both chromosomes and plasmids, of epidemic C. difficile ST37 strains in China.

ABO blood group-associated susceptibility to norovirus infection: A systematic review and meta-analysis.

BACKGROUND: Norovirus is responsible for the viral gastroenteritis burden worldwide. Histo-blood type antigens (HBGAs) are the only well-known factor regarding their effect on the pathogenesis of norovirus. Here, we performed the study to further investigate the association of the ABO blood group with norovirus susceptibility. METHODS: All relevant studies were retrieved from PubMed, Embase, Web of Science, and Cochrane Library databases and the associations of ABO blood groups with norovirus were assessed. The pooled odds ratios (ORs) and 95% confidence interval (CI) were calculated from extracted data. I2 statistics, sensitivity analysis, and publication bias were used to confirm the findings. Subgroup analyses were performed for genotypes, publication years, development degree of the countries, and age if heterogeneity was recorded. RESULTS: Seventeen articles covering 2304 participants were included. The overall analysis of the studies showed similar ORs of norovirus infection among individuals with blood type A (OR = 0.90, 95%CI = 0.71-1.14, P = 0.37) and blood type B (OR = 0.85, 95%CI = 0.66-1.12, P = 0.25) as compared to those controls. An increased odds of norovirus infection was found among individuals with blood type O (OR = 1.28, 95%CI = 1.03-1.59, P = 0.03), while the individuals with blood type AB (OR = 0.91, 95%CI = 0.60-1.39, P = 0.67) showed no correlation with norovirus infection. For blood type B and blood type AB, the results of subgroup analyses mirrored the observations above. CONCLUSIONS: The meta-analysis suggested that the blood type A, B and AB might not affect susceptibility to norovirus infection. However, blood type O appeared to be more susceptible to norovirus infection.

Diagnostic test accuracy of droplet digital PCR for the detection of EGFR mutation (T790M) in plasma: Systematic review and meta-analysis.

BACKGROUND: T790M mutation was a primary lead cause in the acquired resistance to EGFR-TKIs confirmed in earlier studies. Since the shortcomings of tumor tissue detection are well known, the liquid biopsy is more appropriate to track T790M status. We assessed the accuracy and clinical significance of the droplet digital PCR (ddPCR) detection of T790M mutation in plasma. METHODS: We retrieved PubMed, Embase, Cochrane, and Web of science with no limitation of language and publication year. Summary sensitivity and specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR) and diagnostic odds ratio of detection of EGFR T790M status were calculated from extracted data from included articles. The summary receiver operating curve (SROC), diagnostic odds ratio (DOR), and the area under the summary receiver operating curve (AUC) was used to assess the overall diagnostic accuracy. I2 and meta-regression were used to evaluate heterogeneity and the source of heterogeneity, respectively. RESULT: We identified 15 studies in the total search of 1364 reports, including 427 paired tissue and plasma samples. The pooled sensitivity and the pooled specificity were 0.68 (95% CI 0.61-0.75) and 0.85 (95% CI 0.75-0.91) by the bivariate model, respectively. The AUC and the pooled DOR were 0.78 (95% CI 0.74-0.81) and 12 (95% CI 7-22), respectively. None of the cofactors could account for the heterogeneity. CONCLUSION: The plasma analysis is of a promising performance to screen EGFR-T790M mutation status by ddPCR.

HLA-A, -B, -DRB1 Alleles as Genetic Predictive Factors for Dengue Disease: A Systematic Review and Meta-Analysis.

Dengue virus infection (DEN) is one of the most prevalent arbovirus diseases in the tropical and subtropical areas. Some human leukocyte antigen (HLA) alleles have been reported to be a protective or risk factor to DEN. Due to the limited sample sizes and regional limitations, the results of individual studies were various. This meta-analysis aimed at investigating the relationship between HLA alleles and dengue disease. Relevant studies of the relationship between HLA and dengue disease were searched through PubMed, Embase, Web of science, and Cochrane databases. Subgroups according to ethnicity or sub-alleles and sensitivity analysis were used to explore the potential source of heterogeneity, which was performed to confirm the findings. The relationships between HLA and dengue disease were defined by odds ratios (ORs) with a 95% confidence interval (CI). Fourteen studies were finally confirmed. Results indicated that A*0203 (OR = 2.19, 95% CI = 1.30-3.69) and A*24 in the Asian group (OR = 1.44, 95% CI = 1.21-1.71) were positively associated with an increased risk of DEN when compared with normal controls. A*33 (OR = 0.49, 95% CI = 0.34-0.69) in Southeast Asia was negatively associated with DEN when compared with normal controls, suggesting a protective role against DEN. In addition, DRB1*11 (OR = 4.10, 95% CI = 1.23-13.69) was positively associated with severe dengue (SD) when compared with dengue fever, whereas DRB1*03 (OR = 0.48, 95% CI = 0.28-0.82) and DRB1*09 (OR = 0.73, 95% CI = 0.55-0.96) were negatively associated with SD when compared with normal controls. The meta-analysis confirmed that HLA-A*0203, A*24, A*33, DRB1*03, DRB1*09, and DRB1*11 have significantly affected dengue disease, and the associations are related to race and regions.

Transcriptomics Study on Staphylococcus aureus Biofilm Under Low Concentration of Ampicillin.

Staphylococcus aureus is one of the representative foodborne pathogens which forms biofilm. Antibiotics are widely applied in livestock husbandry to maintain animal health and productivity, thus contribute to the dissemination of antimicrobial resistant livestock and human pathogens, and pose a significant public health threat. Effect of antibiotic pressure on S. aureus biofilm formation, as well as the mechanism, remains unclear. In this study, the regulatory mechanism of low concentration of ampicillin on S. aureus biofilm formation was elucidated. The viability and biomass of biofilm with and without 1/4 MIC ampicillin treatment for 8 h were determined by XTT and crystal violet straining assays, respectively. Transcriptomics analysis on ampicillin-induced and non-ampicillin-induced biofilms were performed by RNA-sequencing, differentially expressed genes identification and annotation, GO functional and KEGG pathway enrichment. The viability and biomass of ampicillin-induced biofilm showed dramatical increase compared to the non-ampicillin-induced biofilm. A total of 530 differentially expressed genes (DEGs) with 167 and 363 genes showing up- and down-regulation, respectively, were obtained. Upon GO functional enrichment, 183, 252, and 21 specific GO terms in biological process, molecular function and cellular component were identified, respectively. Eight KEGG pathways including "Microbial metabolism in diverse environments", "S. aureus infection", and "Monobactam biosynthesis" were significantly enriched. In addition, "beta-lactam resistance" pathway was also highly enriched. In ampicillin-induced biofilm, the significant up-regulation of genes encoding multidrug resistance efflux pump AbcA, penicillin binding proteins PBP1, PBP1a/2, and PBP3, and antimicrobial resistance proteins VraF, VraG, Dlt, and Aur indicated the positive response of S. aureus to ampicillin. The up-regulation of genes encoding surface proteins ClfB, IsdA, and SasG and genes (cap5B and cap5C) which promote the adhesion of S. aureus in ampicillin induced biofilm might explain the enhanced biofilm viability and biomass.

Whole-genome sequencing identification of a multidrug-resistant Salmonella enterica serovar Typhimurium strain carrying blaNDM-5 from Guangdong, China.

A carbapenem-resistant Salmonella enterica serovar Typhimurium (sequence type 34 [ST34]) strain was isolated from a fecal specimen from a child with acute diarrhea. Whole-genome sequencing revealed that the 84.5-kb IncFII plasmid pST41-NDM carrying the NDM-5 carbapenemase gene possesses a structure identical to that of the IncFII-type plasmid backbone. However, the blaNDM-5 flanking sequence found in this plasmid is identical to the blaNDM-5-positive IncX3 plasmids carried by 10 strains of Enterobacteriaceae identified in the same hospital.

Emergence and Plasmid Analysis of Klebsiella pneumoniae KP01 Carrying blaGES-5 from Guangzhou, China.

Klebsiella pneumoniae strain KP01 carrying blaGES-5 was identified from a patient in Guangzhou, China. High-throughput sequencing assigned blaGES-5 to a 28.5-kb nonconjugative plasmid, pGES-GZ. A 13-kb plasmid backbone sequence on pGES-GZ was found to share high sequence identities with plasmids from Gram-negative nonfermenters. A novel class 1 integron carrying a gene cassette array of orf28-orf28-blaGES-5 was identified on pGES-GZ, within which orf28 encoded a hypothetical protein possibly correlated to fosfomycin resistance.

resistome analysis of Enterobacter cloacae CY01, an extensively drug-resistant strain producing VIM-1 metallo-ß-lactamase from China.

Resistome analysis of clinical VIM-1-producing Enterobacter cloacae strain CY01 from China revealed the presence of multiple resistance determinants. Two resistance plasmids were identified in CY01. The pCY-VIM plasmid was 14 kb in size and possessed a replicase gene (repA), a gene cluster encoding the partitioning function (parABC), and a carbapenemase gene (blaVIM-1). Another 5.9-kb plasmid, pCY-MdT, with an aac(6')-Ib gene, was very closely related (13 nucleotide differences) to pMdT1, a ColE1 plasmid carrying aac(6')-Ib-cr4.

Prevalence and characterization of quinolone resistance in Laribacter hongkongensis from grass carp and Chinese tiger frog.

Laribacter hongkongensis is a food-borne bacterium associated with community-acquired gastroenteritis and diarrhoea. Quinolone resistance was recently reported in bacterial isolates from aquatic products, but the molecular mechanisms for resistance were still unknown. In this study, a total of 157 L. hongkongensis strains were isolated from grass carps (n = 443) and Chinese tiger frogs (n = 171). Twenty-one ciprofloxacin-resistant strains were analysed for mutations in quinolone resistance-determining regions (QRDR), acquired quinolone resistance (AQR) genes and the role of efflux pumps in resistance. All QRDR mutations in gyrA (codons 85 and 89) and parC (codons 83 and 231) were found to be closely associated with ciprofloxacin resistance. The AQR gene aac(6')-Ib-cr was found in 42.9% (9/21) of the resistant strains, but qnrA, qnrB, qnrC, qnrD, qnrS and qepA were not detected. No significant change of MICs to ciprofloxacin was observed in the presence of an efflux pump inhibitor, indicating the role of efflux pump was probably absent. All 21 ciprofloxacin-resistant strains showed different electrophoretic patterns, which suggested they were not genetically related. These data highlight the importance of QRDR mutations and the AQR gene aac(6')-Ib-cr during the development of quinolone resistance in a heterogeneous population of L. hongkongensis.

[Evaluation of the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis].

OBJECTIVE: To evaluate the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis. METHODS: A total of 121 patients with osteoarticular tuberculosis, who were hospitalized in Shenzhen Third People's Hospital during June 2008 to June 2012, were recruited for analysis. Clinical laboratory tests results were collected for comparison between patients with or without chronic co-infection with virus. RESULTS: Among the 121 patients, thirty patients were co-infected with hepatitis B virus (HBV), two were with Human immunodeficiency virus (HIV), and one was co-infected with HBV, HIV and hepatitis C virus (HCV). Compared to patients with osteoarticular tuberculosis without HBV/HCV/HIV infection, patients with chronic HBV/HCV/HIV virus infection had similar positive rate of laboratory tests including tissue smear acid-fast bacilli (AFB) staining, tissue Mycobacterium tuberculosis (Mtb) culture, tissue Mtb DNA detection, serological test of antibodies against Mtb, and Mtb. antigen-specific interferon-gamma release assay. Similar results were also found for erythrocyte sedimentation rate, C-reative protein level and liver function including Alanine aminotransferase and Aspartate Aminotransferase. CONCLUSION: Chronic infection with HBV/HCV in patients with have no obvious effect on clinical laboratory tests related to tuberculosis.