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Rongrong Li,1,2 Huaming Xu,3 Hao Tang,4 Jilu Shen,1,5 Yuanhong Xu1 1Department of Clinical Laboratory, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People’s Republic of China; 2Department of Pathogen Biology and Provincial Laboratories of Pathogen Biology and Zoonoses, Anhui Medical University, Hefei, People’s Republic of China; 3The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei, People’s Republic of China; 4Department of Clinical Laboratory, The Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 5Anhui Public Health Clinical Center, Hefei, People’s Republic of ChinaCorrespondence: Yuanhong Xu, Department of Clinical Laboratory, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People’s Republic of China, Tel +86 13505694447, Email xyhong1964@163.comBackground: Bloodstream infection (BSI) is a common type of infection frequently diagnosed in clinics. The emergence and spread of ESBLs-producing Escherichia coli (E. coli) has emerged as one of the biggest challenges in global community health.Methods: The production of ESBLs was determined by the composite disk diffusion method. The expression of the various resistance and virulence genes were detected by PCR and sequencing. Multi-locus sequence typing (MLST) and phylogenetic groups were used for the classification. The transfer of resistant plasmids was determined by conjugation assay. The statistical differences were analyzed using Statistical Product and Service Solutions (SPSS) version 23.0.Results: A total of 60 strains of ESBLs-producing E. coli were collected. The resistance genes that were identified included blaCTX-M, blaTEM, blaSHV, blaOXA-1 and mcr-1. The most common one was the blaCTX-M including blaCTX-M-27 (n = 16), blaCTX-M-14 (n = 15), blaCTX-M-15 (n = 11), blaCTX-M-55 (n = 14) and blaCTX-M-65 (n = 5). A total of 31 STs were detected, and the most abundant amo

Yi Zhu,1,2 Chunhui Lin,1,2 Huaming Xu,3 Zhaoxin Xia,1,2 Wensu Yang,1,2 Hao Tang,4 Xinyi Hu,1,2 Tong Jiang,1,2 Zhen Liu,1,2 Jilu Shen1,2 1The First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 2Anhui Public Health Clinical Center, Hefei, People’s Republic of China; 3The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, People’s Republic of China; 4The Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of ChinaCorrespondence: Jilu Shen, Tel +86 151 5515 2963, Email shenjilu@ahmu.edu.cn; 1464675852@qq.comIntroduction: More than half of the world’s people are infected or have been infected with Helicobacter pylori. This infection is related to many diseases, with its pathogenicity related to virulence factors. Therefore, the rapid diagnosis of H. pylori and genotyping of virulence genes play an extremely important role in the clinical treatment and control of transmission.Methods: To this end, we developed a molecular detection method based on RPA- CRISPR-Cas12a technology for the specific genes 16S rDNA gene, cytotoxin associated gene A(cagA), and vacuolating cytotoxin A (vacA) of H. pylori.Results: The results of which were displayed by lateral flow strips. Macroscopic observation takes only about 25 minutes and the sensitivity is 2ng/microliter.Discussion: The method is simple, convenient to operate and has low costs, and can therefore be applied widely to the detection and typing of H. pylori in various environments such as primary hospitals, community clinics, outdoors, and large medical institutions.Keywords: Helicobacter pylori, virulence genes, recombinase polymerase amplification, CRISPR-Cas12a, lateral flow immunochromatographic strip

Chunhui Lin,1,2,* Hao Tang,3,* Xinyi Hu,1,2 Ge Li,1,2 Tong Jiang,1,2 Wensu Yang,1,2 Zhaoxin Xia,1,2 Yi Zhu,1,2 Huaming Xu,4 Jing Zhou,1,2 Jilu Shen1,2 1Clinical Laboratory, the First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 2Clinical Laboratory, Anhui Public Health Clinical Center Hefei, Hefei, People’s Republic of China; 3Clinical Laboratory, the Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 4Clinical Laboratory, the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jilu Shen, Tel +86 151 5515 2963, Email shenjilu@ahmu.edu.cnIntroduction: It is time-consuming to identify fungal pathogens from positive blood cultures using the standard culture-based method. And delayed diagnosis of bloodstream infection leads to significantly increased mortality.Methods: We developed a PCR-reverse dot blot hybridization combined with microfluidic chip techniques to rapidly identify 13 fungal pathogens within 3– 4 h using the sample of blood cultured over a period of time.Results: We performed clinical validation using 43 blood culture-positive samples with a sensitivity of 96.7%, a specificity of 100%, and a concordance rate of 97.7%. Samples with different culture durations were evaluated using our approach, showing a detection rate of 85.2% at 16 h and 96.3% at 24 h; the platform could reach a detection limit of 103cfu/mL for the Candida spp. and 103 copies/mL for Aspergillus spp.Discussion: The detection rate of the platform is much higher than the positive rates of concurrent blood cultures. This method bears substantial clinical application potential as it incorporates the microfluidic platform with low reagent consumption, automation, and low cost (about 10 dollars).Keywords: fungemia, bloodstream infection, microfluidics, Candida aur

Wensu Yang,1,2 Zhaoxin Xia,1,2 Yi Zhu,1,2 Hao Tang,1,2 Huaming Xu,1,2 Xinyi Hu,1,2 Chunhui Lin,1,2 Tong Jiang,1,2 Pei He,1,2 Jilu Shen1,2 1The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 23001, People’s Republic of China; 2Anhui Public Health Clinical Center Hefei, Anhui, 230012, People’s Republic of ChinaCorrespondence: Jilu Shen, Tel +86-15155152963, Email shenjilu@ahmu.edu.cnBackground: Fecal microbiota transplantation (FMT) has emerged as a new therapy targeting gastrointestinal microbiota for the treatment of a growing number of diseases in recent years. Previous studies have suggested that FMT may be a potential therapy for type 2 diabetes (T2D), but the underlying mechanism remains unclear. Therefore, in the present study, we aimed to investigate the role of FMT in T2D and its underlying mechanisms.Methods: To induce T2D, mice were fed a high-fat diet and injected with low-dose streptozotocin (STZ) for four weeks. The mice were then randomly divided into four groups: control group (n = 7), T2D group (n = 7), metformin (MET)-treated group (n = 7), and FMT group (n = 7). The MET group was orally administered 0.2 g/kg MET, the FMT group was orally administered 0.3 mL of bacterial solution, and the other two groups were orally administered the same volume of saline for four weeks. Serum and fecal samples were collected for non-targeted metabolomics, biochemical indicators, and 16S rRNA sequencing, respectively.Results: Our results demonstrated that FMT had a curative effect on T2D by ameliorating hyperlipidemia and hyperglycemia. Using 16S rRNA sequencing and serum untargeted metabolomic analysis, we found that FMT could restore the disorders of gastrointestinal microbiota in T2D mice. Moreover, corticosterone, progesterone, L-urobilin, and other molecules were identified as biomarkers after FMT treatment. Our bioinformatics analysis suggested that steroid hormone biosynthesis, arginine, proline metabolism, and unsaturated fatty acid

Pei He,1,2 Jie Xia,3 Peixin Zhang,3 Wensu Yang,1,2 Zhaoxin Xia,1,2 Ping Liu,4 Yi Zhu,1,2 Yaping Fang,5 Zhenhua Zhang,3 Jilu Shen1,2 1Department of Laboratory Medicine, The First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 2Department of Laboratory Medicine, Anhui Public Health Clinical Center, Hefei, People’s Republic of China; 3Department of Infectious Diseases, The Second Hospital of Anhui Medical University, Hefei, People’s Republic of China; 4Department of Infectious Diseases, People’s Hospital of Jieshou City, Fuyang, People’s Republic of China; 5Department of Clinical Laboratory, The Second Hospital of Anhui Medical University, Hefei, People’s Republic of ChinaCorrespondence: Zhenhua Zhang; Jilu Shen, Email zzh1974cn@163.com; shenjilu@ahmu.edu.cnAim: Hepatitis B surface antibody (HBsAb) plays an important role in the prevention of hepatitis B virus (HBV) infection, especially in immunocompromised individuals and in those infected with HBV.HBsAb levels often fluctuate and decrease.This study aimed to determine the regularity of HBsAb persistence among different populations. Moreover, the risk factors and the optimal cutoff value were determined to predict a decreasing population in HBsAb level.Methods: The study involved 182 participants, including 76 patients with a 25% decrease in HBsAb levels and 106 patients with an HBsAb decrease rate of > 50%. Both hepatitis B core antibody negative and positive patients were included.These patients were followed up for 10 years. The follow-up demographic and laboratory data were recorded and compared among the groups. Fluctuations in HBsAb data and HBsAb persistent immunity were evaluated. The independent factors and the optimal cutoff value were recorded.Results: The first HBsAb median of Group 4 was lower than that of the other groups, and its median was 50.8 mlU/mL. In addition, the persistent immunity of the case groups was shorter than that of the control groups

Guoping Lu,1,2 Hao Tang,3 Zhaoxin Xia,1 Wensu Yang,1 Huaming Xu,1 Zhen Liu,1 Shenwang Ni,1 Zhaofei Wang,1 Jilu Shen1 1Department of Laboratory Medicine, The First Affiliated Hospital of Anhui Medical University; Anhui Public Health Clinical Center, Hefei, People’s Republic of China; 2Department of Laboratory Medicine, The Affiliated Fuyang Hospital of Anhui Medical University, Fuyang, People’s Republic of China; 3Department of Laboratory Medicine, The Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of ChinaCorrespondence: Jilu Shen, The First Affiliated Hospital of Anhui Medical University; Anhui Public Health Clinical Center, Hefei, People’s Republic of China, Email shenjilu@ahmu.edu.cnIntroduction: To examine the in vitro and in vivo antimicrobial activities of ceftazidime/avibactam (CZA) alone or in combination with aztreonam (ATM) against KPC-, NDM-, IMP-, KPC+IMP-, KPC+NDM-producing strains.Methods: A total of 67 clinical non-repetitive carbapenem-resistant Enterobacterales (CRE) strains were selected for the microdilution broth method that was performed to analyze the minimal inhibitory concentration (MIC) and the combination antimicrobial susceptibility test using checkerboard titration method. The fractional inhibitory concentration (FIC) was calculated to determine the antimicrobial effect. The time-kill assays and the mouse infection model were used to study the bactericidal effect and therapeutic effect of CZA alone or in combination with ATM.Results: The CZA minimal inhibitory concentration (MIC) values of CZA revealed that 29 KPC-producing strains and 1 OXA-producing strain were ≤ 4μg/mL. The CZA MIC values of 37 metal-β-lactamase (MBLs)-producing strains such as NDM-, IMP-, KPC+IMP-, KPC+NDM-producing strains were ≥ 128μg/mL, after combining with ATM, the FIC values were all below 0.51. The time-kill assays revealed that CZA at various concentrations of 2, 4 and 8 MIC showed significant bactericidal effic

Huaming Xu,1 Hao Tang,2 Rongrong Li,3 Zhaoxin Xia,2 Wensu Yang,2 Yi Zhu,2 Zhen Liu,2 Guoping Lu,4 Shenwang Ni,2 Jilu Shen1 1Department of Laboratory Medicine, The Fourth Affiliated Hospital of Anhui Medical University, Hefei, 230012, People’s Republic of China; 2University Laboratory, The Fourth Affiliated Hospital of Anhui Medical University, Hefei, 230012, People’s Republic of China; 3The First Affiliated Hospital of Anhui Medical University Laboratory Department, Hefei, Anhui, Peoples’ Republic of China; 4Laboratory Department of Fuyang Hospital Affiliated to Anhui Medical University, Fuyang, Anhui, People’s Republic of ChinaCorrespondence: Jilu Shen, Tel +86 151 5515 2963, Email shenjilu@ahmu.edu.cn; 1464675852@qq.comIntroduction: Carbapenemase-mediated antimicrobial resistance is currently a hot spot of global concern. Carbapenem-resistant organisms are highly prevalent in hospitals associated with difficult-to-treat infections, resulting in poor clinical outcome due to limited treatment options. It is urgently needed to have a rapid, efficient, and convenient molecular assay for identifying such resistant strains.Methods: For this end, we developed a new laboratory assay targeting Klebsiella pneumoniae carbapenemase (KPC) and New Delhi metallo-β-lactamase (NDM) based on loop-mediated isothermal amplification, CRISPR–Cas12a, and lateral flow immunochromatographic strip (CRISPR–Cas-LAMP-lateral flow strip). The method was designed to use a guide RNA (gRNA) to recognize the target DNA and guide Cas12a to cleave the target DNA, and simultaneously cleave any single-stranded DNA within the cleavage reaction system.Results: The cleavage products are visible to the naked eye on the lateral flow strip. This method is highly sensitive in direct detection of bacteria in samples containing at least 3× 105 CFU/mL without the need for bacterial culture.Discussion: It provides shorter turnaround time and higher specificity than the conventional bacterial cultur

Zhen Liu,1 Hao Tang,1 Huaming Xu,1 Guoping Lu,2 Wensu Yang,1 Zhaoxin Xia,1 Yi Zhu,1 Shenwang Ni,1 Wanqi Men,1 Jilu Shen1 1The Fourth Affiliated Hospital of Anhui Medical University Laboratory Department, Hefei, People’s Republic of China; 2Laboratory Department of Fuyang Hospital Affiliated to Anhui Medical University, Fuyang, Anhui, 236000, People’s Republic of ChinaCorrespondence: Jilu Shen, The Fourth Affiliated Hospital of Anhui Medical University Laboratory, No. 100 Huaihai Avenue, Xinzhan District, Hefei, Anhui Province, 230012, People’s Republic of China, Tel +86 151 5515 2963, Email shenjilu@ahmu.edu.cnAim: To reduce the inspection time for urinary tract pathogens and provide a rapid and effective therapeutic plan for clinical anti-infection treatment, this study developed a rapid identification (ID) and antimicrobial sensitivity test (AST) method by DOT-MGA.Methods: We grouped midstream urine samples with single bacteria according to the number of bacteria (≤ 5/5– 15/≥ 15) under per oil microscope after Gram staining. Then we adopted differential centrifugation to process the grouped samples to collect precipitate. MALDI-TOF MS was performed using precipitate directly or after short-term culture. If succeed, we resuspended the precipitate into droplets with or without antibiotics at a MALDI target. Four hours later, mass spectrometer (MS) was used to identify the culture on the target and to analyse AST.Results: Samples (count ≥ 15), which precipitate can be directly identified by MS; otherwise, the precipitate need a short-term cultured for 3– 6 h before ID. The consistency of the ID results between conventional culture and the precipitate is 100%. Compared with broth microdilution method, DOT-MGA for predicting AST had a high consistency. EA and CA for IPM, LEV, CAZ, NIT, and FOT were 100%/100%, 98%/90%, 98%/92%, 100%/90%, 98%/94%, respectively. No VME was observed in all tests. Besides, MIC50 for the five antibiotics by DOT-MGA and broth

Hao Tang,1 Rongrong Li,2 Huaming Xu,1 Guoping Lu,3 Zhen Liu,1 Wensu Yang,1 Zhaoxin Xia,1 Yi Zhu,1 Jilu Shen1 1The Fourth Affiliated Hospital of Anhui Medical University Laboratory Department, Hefei, People’s Republic of China; 2The First Affiliated Hospital of Anhui Medical University Laboratory Department, Hefei, People’s Republic of China; 3Laboratory Department of Fuyang Hospital Affiliated to Anhui Medical University, Fuyang, Anhui, 236000, People’s Republic of ChinaCorrespondence: Jilu ShenThe Fourth Affiliated Hospital of Anhui Medical University Laboratory, No. 100 Huaihai Avenue, Xinzhan District, Hefei, Anhui Province, 230012, People’s Republic of ChinaTel +86 151 5515 2963Email shenjilu@ahmu.edu.cnIntroduction: The recently developed DOT-MGA (direct-on-target microdroplet growth assay) has shown the desirability of direct application of this approach in positive blood cultures and its good performance in detection. This study selected 44 Enterobacteriaceae strains and implemented a DOT-MGA assay on blood cultures to detect their resistance to seven antibiotics. The results of DOT-MGA were compared with the other two antimicrobial susceptibility testing (AST) methods to analyze the detection performance of DOT-MGA.Methods: We adopted the differential centrifugation to process positive blood-culture (BC). Processed BC broth was directly used for rapid AST using DOT-MGA. Droplets of 6 μL with and without antibiotics at the EUCAST breakpoint concentration were spotted in triplicates onto the surface of a MALDI target. The plates were incubated in a wet box for 4 h before the broth was removed with filter paper. Bruker Biotyper software was used to analyze the test results compared with standard database, and the scores were used to quantify and determine the results.Results: DOT-MGA results were compared with the direct-from-BC disk-diffusion method and results were reported by broth microdilution method, respectively. The comparison demonstrated a 10