scholarly journals TaqMan Array card testing of participant-collected stool smears to determine the pathogen-specific epidemiology of travelers’ diarrhea

2021 ◽  
Author(s):  
Michele D Tisdale ◽  
David R Tribble ◽  
Indrani Mitra ◽  
Kalyani Telu ◽  
Huai-Ching Kuo ◽  
...  
Keyword(s):  
Loose Stool ◽  
Storage Duration ◽  
E Coli ◽  
Fta Cards ◽  
Common Pathogen ◽  
Qualitative Detection ◽  
Diarrheal Episode ◽  
Improve Compliance ◽  
Observational Cohort ◽  
Taqman Array

Abstract Background: We assessed the compliance with self-collection of stool smears on Whatman® FTA® Elute Card (FTA Card) and detection of travelers’ diarrhea (TD) associated pathogens using a quantitative PCR assay (customized TaqMan® array card [TAC]), in a prospective, observational cohort of travelers. Methods: Enrolled travelers documented symptoms on a travel diary and collected an FTA Card during a diarrheal episode, or at the end of travel if they remained asymptomatic. TAC testing was performed on FTA Cards from TD cases and 1:1 matched asymptomatic controls and 1:1 matched loose stool cases that did not meet TD criteria. Odds ratios (OR) were used to determine the association between detected pathogens and TD. Results: 484 of 2456 (19.7%) travelers completed an illness diary and met TD criteria, and 257 (53.1%) collected an FTA Card during the TD episode. FTA Cards were stored for a median of 2 years at room temperature (IQR: 1-4 years) before extraction and testing. The overall TAC detection rate in TD cases was 58.8% (95%CI: 52.5-64.8). Enterotoxigenic E. coli was the most common pathogen in TD cases (26.8%) and 3.5% of samples were positive for norovirus. The odds of detecting TD-associated pathogens in 231 matched cases and asymptomatic controls was 5.4 (95% CI: 3.6-8.1) and 2.0 (95% CI:1.1-3.7) in 121 matched TD and loose stool cases (p < 0.05). Enteroaggregative E coli was the most common pathogen detected in asymptomatic controls and loose stool cases. Detection of diarrheagenic E coli, Shigella/enteroinvasive E coli (EIEC), and Campylobacter spp. was significantly associated with TD. Conclusions: FTA Cards are a useful adjunct to traditional stool collection methods for evaluating the pathogen-specific epidemiology of TD in austere environments. Qualitative detection of pathogens was associated with TD. Measures to improve compliance and quality of FTA Card collection with decreased storage duration may further optimize detection.

scholarly journals DiarrheagenicEscherichia coliPhylogroups Are Associated with Antibiotic Resistance and Duration of Diarrheal Episode

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Susan Mosquito ◽  
Maria J. Pons ◽  
Maribel Riveros ◽  
Joaquim Ruiz ◽  
Theresa J. Ochoa
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Clinical Characteristics ◽  
Healthy Controls ◽  
Persistent Diarrhea ◽  
Phylogenetic Groups ◽  
E Coli ◽  
Diarrheal Episode ◽  
Group A ◽  
Group D

Conventionally, inEscherichia coli, phylogenetic groups A and B1 are associated with commensal strains while B2 and D are associated with extraintestinal strains. The aim of this study was to evaluate diarrheagenic (DEC) and commensalE. coliphylogeny and its association with antibiotic resistance and clinical characteristics of the diarrheal episode. Phylogenetic groups and antibiotic resistance of 369E. colistrains (commensal strains and DEC from children with or without diarrhea) isolated from Peruvian children <1 year of age were determined by a Clermont triplex PCR and Kirby-Bauer method, respectively. The distribution of the 369E. colistrains among the 4 phylogenetic groups was A (40%), D (31%), B1 (21%), and B2 (8%). DEC-control strains were more associated with group A while DEC-diarrhea strains were more associated with group D(P<0.05). There was a tendency(P=0.06)for higher proportion of persistent diarrhea (≥14 days) among severe groups (B2 and D) in comparison with nonsevere groups (A and B1). Strains belonging to group D presented significantly higher percentages of multidrug resistance than the rest of the groups(P>0.01). In summary, DEC-diarrhea strains were more associated with group D than strains from healthy controls.

scholarly journals Microbial occurrence at different freezing duration in speckled shrimp, Metapenaeus monoceros (Fabricius, 1798) from local markets of Dhaka city

2013 ◽  
Vol 22 (2) ◽  
pp. 91-101
Author(s):  
Md Anisur Rahman Khan ◽  
Tamanna Yasmin ◽  
Saima Sharif Nilla ◽  
Md Mahmudur Rahman Khan ◽  
Naziza Rahman ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Total Coliform ◽  
Microbial Load ◽  
Dhaka City ◽  
Storage Duration ◽  
Local Markets ◽  
E Coli ◽  
Coliform Count ◽  
Metapenaeus Monoceros ◽  
Significant Difference

A study was carried out at four different (0, 1, 7 and 30 days) freezing durations in regard to the microbiological incidence in speckled shrimp (Metapenaeus monoceros) from three different local markets - Ananda bazaar, New Market and Polashi bazaar of Dhaka city. The study unveiled that Ananda bazaar samples were more or less contaminated than those of others. The microbial load varied with market sources as total bacteria count (TBC), total coliform count (TCC) and faecal coliform count (FCC) were found in all the storage periods crossing the acceptable limit. Except for E. coli (EC), all the bacterial counts were significantly different (p < 0.05) in 7 and 30 days freezing durations. A significant difference (p < 0.05) was found in all freezing durations only for EC counts. After 30 days storage duration for all counts showed 101 cfu/g reduction of microbial load from every initial count. The Salmonella-Shigella (SS) and Vibrio spp. were identified in all the samples of the three markets. In case of biochemical composition, an inverse relation was found in moisture - protein and moisture - fat where the moisture content increased after 30 days freezing duration but protein and lipid contents decreased with time. This study revealed that shrimp available in local markets of Dhaka city was highly ingested with pathogenic bacteria which indicate the unhygienic condition of the market premises. Dhaka Univ. J. Biol. Sci. 22(2): 91-101, 2013 (July)

scholarly journals Common Pathogen and their Drug Sensitivity Pattern in Urinary Tract Infection at Chandka Medical College Hospital Larkana

2021 ◽  
Vol 15 (7) ◽  
pp. 1868-1872
Author(s):  
Hakim Ali Abro ◽  
Sheeraz Ali ◽  
Sultan Ahmed Chandio ◽  
Jamil Muqtadir ◽  
Mumtaz Ali Chutto ◽  
...  
Keyword(s):  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Drug Sensitivity ◽  
University Hospital ◽  
Causative Organism ◽  
Klebsiella Pneumonia ◽  
E Coli ◽  
Common Pathogen ◽  
Sensitivity Pattern

Objective: To study the pattern of existing common pathogen of Urinary Tract Infection and their drug sensitivity pattern. Methods: It was a cross sectional study held in Medical Unit-I, CMC Teaching Hospital, Shaheed Mohtarma Benazir Bhutto Medical University(SMBBMU), Larkana with collaboration of Pathology Laboratory CMC Larkana, Aga Khan University Hospital Lab, Rahila Research Lab and Chughtai Lab. All the participants meeting inclusion criteria were enrolled. The reports of Urine culture were collected from above mentioned laboratories which include bio data, causative organism, their count and drug sensitivity pattern. The results were copied on proforma by researcher himself. SPSS version 25 was applied for data analysis. Results: Total 302 participants were registered in the study. Out of them 75.8% of them were female and 24.2% were male. Mean age of the patients was 40 ± 9 years. E-coli was found in 52.3%, Enterococcus in 13.2% and Klebsiella pneumonia in 9.6% cases. Meropenem was found sensitive in 92.4%, while cefoperazone sulbactam, Amikacin and Fosfomycin were found to be sensitive in 86.6%, 85%, 76.2% respectively. Ampicillin and Vancomycin was used in 50 cases for Enterococci. Ampicillin was found sensitive in 70% of cases where as vancomycin was found sensitive in 100% of cases with no vancomycin resistant Enterococci. For Enterobacteria Moxifloxacin was found resistant in 84.4% of cases, Ampicillin in 83.5%, Nalidix acid in 78% cases. While Ceftazidime and Amoxiclav was found resistant in 71.1% and 54.1% respectively. Conclusion: E-coli, Enterococcus and klabsella were the most pathogens found in this study. The drug sensitivity pattern showed that Meropenem, Cefoperazone Sulbactam, Amikacin, Fosfomycin and Vancomycin were found commonly sensitive while Moxifloxacin, Ampicillin, Ceftazidime and Amoxiclav was found resistant. Keywords: Urinary Tract Infection, Culture and Sensitivity, Meropenem, Vancomycin, Cefoperazone Sulbactam CMC @ SMBBMU, Larkana.

scholarly journals Avaliação das condições higiênico-sanitárias em embarcações mistas da região Amazônica

2021 ◽  
pp. 123-129
Author(s):  
Ariane Kluczkovski ◽  
◽  
Samir Pinto ◽  
Augusto Kluczkovski-Junior ◽  
Marco Pinto ◽  
...  
Keyword(s):  
Food Preparation ◽  
Environmental Evaluation ◽  
Operational Conditions ◽  
E Coli ◽  
Preparation Conditions ◽  
Detection Analysis ◽  
Qualitative Detection ◽  
Sanitary Conditions ◽  
And Training ◽  
Drinking Water Samples

Introduction: The hygienic and sanitary conditions of vessels carrying passengers and cargo requires constant diagnosis and monitoring for disease prevention. Objective: The objective of this study was to identify health hazards in food preparation conditions and water potability in mixed vessels in the State of Amazonas-Brazil. Method: 33 vessels were analyzed with qualitative detection analysis of Escherichia coli by chromogenic substrate for water potability and environmental evaluation with questionnaire application. Results: There was microbiological contamination in 13 (39.39%) drinking water samples and 31 (93.93%) sink samples for E. coli. The structural and operational conditions of kitchen handlers mostly did not meet the minimum percentages of safety in food production. Conclusions: This is the first report on the structural and operational conditions of food preparation in mixed vessels in Manaus-AM and we conclude that intervention is necessary with public policies regarding hygienic and sanitary issues involving the supply of water served in vessels for the protection of passenger health and training of food handlers and inspection of environments.

scholarly journals Burden and Etiology of Moderate and Severe Diarrhea in Chinese Children Less than 5 years of Age: Prospective, Population-based Surveillance

2020 ◽  
Author(s):  
Hong-Lu Zhou ◽  
Theresa Bessey ◽  
Song-Mei Wang ◽  
Zhao-Jun Mo ◽  
Leslie Barclay ◽  
...  
Keyword(s):  
Southern China ◽  
Northern China ◽  
Population Based ◽  
Prevention Measures ◽  
Conventional Pcr ◽  
Effective Prevention ◽  
E Coli ◽  
Severe Diarrhea ◽  
Health Significance ◽  
Taqman Array

Abstract Background. Diarrhea remains the leading cause of childhood illness in China. Better understanding of burden and etiology of diarrheal diseases is important for development of effective prevention measures.Methods. Population-based diarrhea surveillance was conducted in Sanjiang (southern China) year-round and Zhengding (northern China) in autumn/winter. Stool specimens were collected from children <5 years of age experiencing diarrhea. The TaqMan Array Card (TAC), based on multiplex real-time PCR, was applied to detect multiple enteric microbial agents simultaneously. Results using these methods were compared to those derived from conventional PCR assays.Results. During the study period, 6,380 children in Zhengding and 3,581 children in Sanjiang <5 years of age participated. Three hundred and forty (31.2%) and 279 (22.9%) diarrhea episodes were identified as moderate-to-severe in the two counties, with incidence of 60.4 and 88.3 cases per 1,000 child-years in Zhengding and Sanjiang, respectively. The five most frequently detected bacterial and viral agents in Sanjiang were adenovirus, enterovirus, enteroaggregative Escherichia coli (EAEC), rotavirus, and sapovirus all the year round, while the most common viral agents in Zhengding were rotavirus, followed by astrovirus and adenovirus during the cool season. Compared to conventional PCR assay, the average incremental detection via the TAC method was 2-fold.Conclusion. Our study demonstrated high diversity and prevalence of multiple major bacterial and viral agents, including rotavirus and calicivirus, among children in China. Further studies are needed to define the public health significance of neglected but frequently detected pathogens such as EAEC, enterotoxigenic E. coli, Campylobacter, adenovirus, and enterovirus.

scholarly journals Performance Characteristics of the TaqMan Array PCR on Repository Stool Specimens and Smears on Whatman® FTA Elute Cards

2020 ◽  
Author(s):  
Michele Tisdale ◽  
Indrani Mitra ◽  
Andrea J McCoy ◽  
Mark P Simons ◽  
Nathanael D Reynolds ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Quantitative Pcr ◽  
Prolonged Storage ◽  
Storage Duration ◽  
Acid Yield ◽  
Stool Specimens ◽  
Pcr Assays ◽  
Taqman Array ◽  
The Impact ◽  
Archival Specimens

Abstract Objective Stool repositories are a valuable resource for retrospective analyses including quantitative PCR assays to distinguish between asymptomatic shedding and clinical disease. The suitability of archival specimens for this purpose is unclear and requires assessment. We conducted a pilot study to evaluate pathogen detection by TaqMan Array Card (TAC) in travelers’ diarrhea (TD) stool specimens stored for 1–13 years, as well as, the impact of transporting specimens on Whatman FTA Elute cards (FTA Cards) on detection. Results The positive percent agreement (PPA) for TAC on stool vs. microbiologic testing was lower than our a priori PPA estimate of 80% for most pathogens: Shigella spp. (100% [95%CI: 69–100%]), enterotoxigenic E coli (ETEC) (63 [49–75%]), Campylobacter spp. (66 [43–85%]) and Norovirus (37 [16–61%]). Use of the FTA card resulted in a further reduction of PPA. Our findings suggest that archival specimens may lead to insensitive detection on quantitative PCR assays due to degradation of nucleic acid with prolonged storage, although our limited sample size precluded us from evaluating the impact of storage duration on nucleic acid yield. Additional studies are needed to understand the impact of storage duration on quantitative PCR data.

scholarly journals EVALUATION OF ANTIBACTERIAL ACTIVITY OF SOME MEDICINAL PLANTS BY BIOAUTOGRAPHY

2021 ◽  
Author(s):  
Rawaa Al-Kayali ◽  
Joud Jalab ◽  
Adawia Kitaz ◽  
Wassim Abdelwahed
Keyword(s):  
Antibacterial Activity ◽  
Thin Layer ◽  
Peer Review ◽  
Thin Layer Chromatography ◽  
Antibacterial Properties ◽  
Active Compounds ◽  
E Coli ◽  
Acacia Cyanophylla ◽  
Qualitative Detection ◽  
Layer Chromatography

Background Acacia cyanophylla is a medicinal plant of the Fabaceae family that is widely distributed in Australia and Asia, also it has many medicinal properties such as antibacterial and antioxidant activity. Thin layer chromatography (TLC) is wildly used in natural product extract analysis as a finger print. Aim and objective: This study is aimed to conducting a qualitative detection of the active compounds in Acacia cyanophylla, Phlomis syriaca and Scolymus hispanicus plants by thin layer chromatography (TLC) method and studying their antibacterial activity. Methods: the qualitative detection of three plants was conducting using thin layer chromatography (TLC) method. Then, aqueous and ethanolic extracts of the aerial parts of the three plants were extracted using an Ultrasonic bath.  The antibacterial activity on E. coli isolates for six extracts was evaluated using minimum inhibitory concentration (MIC) test. The active compounds that may be responsible for the antibacterial effect was isolated by direct bioautograph method. Results: Performing Thin-layer chromatography TLC tests show  that the three plant contain flavonoids, saponin, bitter principles and essential oils, and all extracts showed antibacterial activity on E. coli isolates, but the ethanolic extract of Acacia cyanophylla was the most effective as the MIC values ranged from 0.097to 3.125mg/mL. Bioautography showed that Escherichia coli was inhibited by most of the separated flavonoids on the TLC plates where four inhibiting spots appeared in yellow color with Acacia cyanophylla and five spots with Scolymus hispanicus, while only one spot appeared with Phlomis syriaca. Conclusion: Acacia cyanophylla extract has been considered as the best antibacterial properties among the selected plants due to the presence of flavonoids                    Peer Review History: Received: 19 July 2021; Revised: 7 August; Accepted: 2 September, Available online: 15 September 2021 Academic Editor:  Ahmad Najib, Universitas Muslim Indonesia, Makassar, Indonesia, [email protected] UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency.  Received file:                Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Sangeetha Arullappan, Universiti Tunku Abdul Rahman, Malaysia, [email protected] Taha A.I. El Bassossy, Medicinal and Aromatic Plants Department, Desert Research Center, Cairo, Egypt, [email protected] Similar Articles: EVALUATION OF ANTIBACTERIAL RESISTANCE OF BIOFILM FORMS OF AVIAN SALMONELLA GALLINARUM TO FLUOROQUINOLONES

scholarly journals The Validation of the VereBeef™ Detection Kit

2019 ◽  
Vol 102 (2) ◽  
pp. 508-524
Author(s):  
Dawn Qian Liu ◽  
Mitsuharu Sato ◽  
Wei Ling Tan ◽  
Grace Pei Wen Khoo ◽  
Kiel Fisher ◽  
...  
Keyword(s):  
False Negative ◽  
Method Comparison ◽  
Department Of Agriculture ◽  
Lab On Chip ◽  
E Coli ◽  
Qualitative Detection ◽  
Assay Performance ◽  
On Chip ◽  
Inspection Service ◽  
Method Comparison Study

Abstract VereBeef™ Detection Kit, incorporating both multiplex PCR and microarray technologies on a lab-on-chip platform, is intended for qualitative detection and differentiation of Escherichia coli O157:H7, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121, E. coli O145, Shiga toxin-producing E. coli (STEC) virulence factors (stx1A, stx2A, eae), and Salmonella species in one test using raw beef trim samples. This product underwent extensive evaluations, including inclusivity-exclusivity, method comparison, robustness, lot-to-lot variability, and stability studies. The inclusivity/exclusivity study demonstrated that VereBeef Detection Kit specifically detects and identifies target analytes without occurrence of false-positive and false-negative detection. In the method comparison study, the performance of the VereBeef Detection Kit was compared with U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook’s methods for target organism detection in raw beef trim using E. coli O157:H7 single inoculation and Salmonella and non-O157 STEC dual inoculation. Data demonstrated equivalence in both methods. The robustness study showed that changesin the test parameters do not impact assay performance. Collectively, VereBeef Detection Kit is able to detect target pathogens in raw beef trim with a minimum enrichment time of 8 h for E. coli O157:H7 detection and 10 h for Salmonella and non-O157 STEC detection.

scholarly journals Burden and etiology of moderate and severe diarrhea in children less than 5 years of age living in north and south of China: Prospective, population-based surveillance

Gut Pathogens ◽  
2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Hong-Lu Zhou ◽  
Theresa Bessey ◽  
Song-Mei Wang ◽  
Zhao-Jun Mo ◽  
Leslie Barclay ◽  
...  
Keyword(s):  
Southern China ◽  
Northern China ◽  
Population Based ◽  
Prevention Measures ◽  
Conventional Pcr ◽  
Effective Prevention ◽  
E Coli ◽  
Severe Diarrhea ◽  
Health Significance ◽  
Taqman Array

Abstract Background Diarrhea remains the leading cause of childhood illness in China. Better understanding of burden and etiology of diarrheal diseases is important for development of effective prevention measures. Methods Population-based diarrhea surveillance was conducted in Sanjiang (southern China) year-round and Zhengding (northern China) in autumn/winter. Stool specimens were collected from children < 5 years of age experiencing diarrhea. The TaqMan Array Card (TAC), based on multiplex real-time PCR, was applied to detect multiple enteric microbial agents simultaneously. Results using these methods were compared to those derived from conventional PCR assays. Results During the study period, 6,380 children in Zhengding and 3,581 children in Sanjiang < 5 years of age participated. Three hundred and forty (31.2%) and 279 (22.9%) diarrhea episodes were identified as moderate-to-severe in the two counties, with incidence of 60.4 and 88.3 cases per 1,000 child-years in Zhengding and Sanjiang, respectively. The five most frequently detected bacterial and viral agents in Sanjiang were adenovirus, enterovirus, enteroaggregative Escherichia coli (EAEC), rotavirus, and sapovirus all the year round, while the most common viral agents in Zhengding were rotavirus, followed by astrovirus and adenovirus during the cool season. Compared to conventional PCR assay, the average incremental detection via the TAC method was twofold. Conclusion Our study demonstrated high diversity and prevalence of multiple major bacterial and viral agents, including rotavirus and calicivirus, among children in China. Further studies are needed to define the public health significance of neglected but frequently detected pathogens such as EAEC, enterotoxigenic E. coli, Campylobacter, adenovirus, and enterovirus.

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