Comparison between Whatman FTA Elute Cards and Conventional Swab for the Detection of Pathogenic Enteric Bacteria Using an RT-qPCR Assay

The emergence of outbreaks of foodborne sickness is carefully related to meals contamination attributable to varied enteric pathogens, resembling Escherichia coli O157:H7, Listeria monocytogenesSalmonella enterica serovar Enteritidis, and Staphylococcus aureus. The management of enteric pathogens poses a problem attributable to the truth that these pathogens can persist for an extended time frame within the surroundings. The fast detection of pathogenic organisms performs a vital function within the prevention and identification of crises associated to well being, security, and well-being. Improper pattern dealing with and processing could affect the diagnostic efficacy and accuracy.
The purpose of the current examine was to check the preservation capability for enteric micro organism between Whatman Flinders Expertise Associates (FTA) playing cards and swabs for reverse transcription-quantitative PCR (RT-qPCR) detection. It was discovered that Whatman FTA playing cards exhibited an improved preservation capability for 5 varieties (each laboratory and environmental strains) of enteric micro organism, together with Escherichia coli O157:H7, Listeria monocytogenesSalmonella enterica serovar Enteritidis, and Staphylococcus aureus for RT-qPCR detection.
Therefore, Whatman FTA playing cards could also be an appropriate device for the routine isolation of foodborne micro organism for molecular prognosis. Subsequently, the usage of Whatman FTA playing cards for pattern assortment and preservation could improve sensitivity and accuracy for micro organism isolation and prognosis.

Analysis of extraction-free RT-qPCR strategies for SARS-CoV-2 diagnostics

Extraction-based real-time reverse transcription quantitative polymerase chain response (RT-qPCR) is at the moment the “gold commonplace” in SARS-CoV-2 diagnostics. Nevertheless, some extraction-free RT-qPCR methods have just lately been developed. On this examine, we in contrast the sensitivity of conventional extraction-based, heated extraction-free, and unheated extraction-free RT-qPCR strategies for SARS-CoV-2 detection in nasopharyngeal swabs from symptomatic people.
The unheated extraction-free methodology confirmed excellent settlement with the usual extraction-based RT-qPCR. In contrast, the heat-treated approach was related to an 8.2% false negativity fee. Unheated extraction-free RT-qPCR for the molecular prognosis of SARS-CoV-2 is a useful various to the standard extraction-based strategies and will speed up turnaround occasions by about two hours.

Speedy dedication of infectious SARS-CoV-2 in PCR-positive samples by SDS-PMA assisted RT-qPCR

The continued COVID-19 pandemic has generated a worldwide well being disaster that wants effectively administration of not solely sufferers but in addition environments to cut back SARS-CoV-2 transmission. The gold commonplace RT-qPCR methodology is delicate and fast to detect SARS-CoV-2 nucleic acid, however doesn’t reply if PCR-positive samples include infectious virions. To avoid this drawback, we report an SDS-propidium monoazide (PMA) assisted RT-qPCR methodology that permits fast discrimination of dwell and lifeless SARS-CoV-2 inside three h. PMA, a photo-reactive dye, can react with viral RNA launched or inside inactivated SARS-CoV-2 virions underneath help of 0.005% SDS, however not viral RNA inside dwell virions. Formation of PMA-RNA conjugates prevents PCR amplification, leaving solely infectious virions to be detected.
Below optimum situations, RT-qPCR detection of heat-inactivated SARS-CoV-2 resulted in bigger than 9 Ct worth variations between PMA-treated and PMA-free teams, whereas lower than 0.5 Ct variations have been noticed within the detection of infectious SARS-CoV-2 starting from 20 to 5148 viral particles. Utilizing a cutoff Ct distinction of 8.6, this methodology might differentiate as little as Eight PFU dwell viruses within the mixtures of dwell and heat-inactivated virions. Additional experiments confirmed that this methodology might efficiently monitor the pure inactivation means of SARS-CoV-2 on plastic surfaces throughout storage with comparable outcomes to the gold commonplace plaque assay. We consider that the culture-free methodology established right here may very well be used for fast and handy dedication of infectious SARS-CoV-2 virions in PCR-positive samples, which can facilitate higher management of SARS-CoV-2 transmission.

The Environmental Microbiology Minimal Info (EMMI) Pointers: qPCR and dPCR High quality and Reporting for Environmental Microbiology


Actual-time quantitative polymerase chain response (qPCR) and digital PCR (dPCR) strategies have revolutionized environmental microbiology, yielding quantitative organism-specific knowledge of nucleic acid targets within the surroundings. Such knowledge are important for characterizing interactions and processes of microbial communities, assessing microbial contaminants within the surroundings (water, air, fomites), and creating interventions (water therapy, floor disinfection, air purification) to curb infectious illness transmission. Nevertheless, our assessment of latest qPCR and dPCR literature in our subject of health-related environmental microbiology confirmed that many researchers should not reporting mandatory and ample controls and strategies, which might serve to strengthen their examine outcomes and conclusions. Right here, we describe the appliance, utility, and interpretation of the suite of controls wanted to make top quality qPCR and dPCR measurements of microorganisms within the surroundings.
Our presentation is organized by the discrete steps and operations typical of this measurement course of. We suggest systematic terminology to reduce ambiguity and assist comparisons amongst research. Instance schemes for batching and mixing controls for environment friendly work movement are demonstrated. We describe important reporting parts for enhancing knowledge credibility, and we offer a component guidelines within the Supporting Info. Moreover, we current a number of key rules in metrology as context for laboratories to plan their very own high quality assurance and high quality management reporting framework.
Following the EMMI tips will enhance comparability and reproducibility amongst qPCR and dPCR research in environmental microbiology, higher inform engineering and public well being actions for stopping illness transmission by way of environmental pathways, and for probably the most urgent points within the self-discipline, focus the load of proof within the route towards options.

CETP Positive Control, Rabbit Serum

KT-151 1 mL
EUR 289

pRedTK-CMV Virus [positive control]

SR10052VA-1 >2 x 10^6 IFUs
EUR 625

pRedZeo-CMV Virus [positive control]

SR10046VA-1 >2 x 10^6 IFUs
EUR 625

Epitope/Affinity Tag Positive Control

BC-144 0.2 mL
EUR 195

pRedTK-CMV Plasmid [positive control]

SR10052PA-1 10 ug
EUR 474

Catenin Antibody (beta + Positive Control)

GWB-T00302 100ug Ask for price

pGreenZeo-CMV Virus [positive control]

SR501VA-1 >2 x 10^6 IFUs
EUR 608

pRedZeo-CMV Plasmid [positive control]

SR10046PA-1 10 ug
EUR 474

pMC.CMV-GFP-SV40PolyA (positive control)

MN601A-1 10 ug
EUR 670

pMC.CMV-rRFP-SV40PolyA (positive control)

MN602A-1 10 ug
EUR 670

pGreenZeo-CMV Plasmid [positive control]

SR501PA-1 10 ug
EUR 474

NATtrol Candida/TV Positive Control (ea)

EUR 446

NATtrol BV Positive Control (6 X 0.15mL)

EUR 446

NATtrol RSV Positive Control (6 x 0.5mL)

NATRSV-6C 6 x 0.5mL
EUR 222

Positive Control Antibody (NTRL-RBD-10)

MAB12443-NTRL-5 0.005
EUR 447.79
Description: Purified Monoclonal Antibody

NATtrol MRSA Positive Control (6 x 0.35mL)

NATMRSA-6L 6 x 0.35mL
EUR 277

NATtrol EV Positive Control (6 X 0.2 mL)

EUR 208

NATtrol SA Positive Control (6 X 0.5 mL)

NATMSSA-6MC 6 X 0.5 mL
EUR 311

NATtrol SA Positive Control  (6 X 0.5 mL)

EUR 331

NATtrol EV Positive Control (6 X 0.5 mL)

MDZ055 6 X 0.2 mL
EUR 208

NATtrol SA Positive Control (6 X 0.5 mL)

MDZ061 6 X 0.5 mL
EUR 378.24
Description: Please contact Gentaur in order to receive the datasheet of the product.

NATtrol RSV Positive Control (6 X 0.5 mL)

NATRSV-6C-IVD 6 X 0.5 mL
EUR 242

NATtrol GBS Positive Control (6 x 0.25 mL)

NATSAG-6L 6 x 0.25 mL
EUR 272
Christopher Miller