Rationally designed upconversion nanoprobe for simultaneous highly sensitive ratiometric detection of fluoride ions and fluorosis theranostics

Biological nanopore-based techniques have attracted more and more attention recently in the field of single-molecule detection, because they allow the real-time, sensitive, high-throughput analysis. Herein, we report an engineered biological...

Download Full-text

Detection by real-time quaking-induced conversion (RT-QuIC), ELISA, and IHC of chronic wasting disease prion in lymph nodes from Pennsylvania white-tailed deer with specific PRNP genotypes

Journal of Veterinary Diagnostic Investigation ◽

10.1177/10406387211021411 ◽

2021 ◽

pp. 104063872110214

Author(s):

Deepanker Tewari ◽

David Steward ◽

Melinda Fasnacht ◽

Julia Livengood

Keyword(s):

Real Time ◽

Disease Susceptibility ◽

Chronic Wasting Disease ◽

Low Frequency ◽

The United States ◽

Detection Methods ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Diagnostic Laboratory ◽

Highly Sensitive

Chronic wasting disease (CWD) is a prion-mediated, transmissible disease of cervids, including deer ( Odocoileus spp.), which is characterized by spongiform encephalopathy and death of the prion-infected animals. Official surveillance in the United States using immunohistochemistry (IHC) and ELISA entails the laborious collection of lymphoid and/or brainstem tissue after death. New, highly sensitive prion detection methods, such as real-time quaking-induced conversion (RT-QuIC), have shown promise in detecting abnormal prions from both antemortem and postmortem specimens. We compared RT-QuIC with ELISA and IHC for CWD detection utilizing deer retropharyngeal lymph node (RLN) tissues in a diagnostic laboratory setting. The RLNs were collected postmortem from hunter-harvested animals. RT-QuIC showed 100% sensitivity and specificity for 50 deer RLN (35 positive by both IHC and ELISA, 15 negative) included in our study. All deer were also genotyped for PRNP polymorphism. Most deer were homozygous at codons 95, 96, 116, and 226 (QQ/GG/AA/QQ genotype, with frequency 0.86), which are the codons implicated in disease susceptibility. Heterozygosity was noticed in Pennsylvania deer, albeit at a very low frequency, for codons 95GS (0.06) and 96QH (0.08), but deer with these genotypes were still found to be CWD prion-infected.

Download Full-text

Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

Virology Journal ◽

10.1186/s12985-020-01467-y ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Yang Zhang ◽

Chunyang Dai ◽

Huiyan Wang ◽

Yong Gao ◽

Tuantuan Li ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Clinical Samples ◽

Chain Reaction ◽

Pcr Assay ◽

Qrt Pcr ◽

Highly Sensitive ◽

One Step ◽

Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

Download Full-text

Probe-Based Real-Time qPCR Assays for a Reliable Differentiation of Capripox Virus Species

Microorganisms ◽

10.3390/microorganisms9040765 ◽

2021 ◽

Vol 9 (4) ◽

pp. 765

Author(s):

Janika Wolff ◽

Martin Beer ◽

Bernd Hoffmann

Keyword(s):

Real Time ◽

Animal Health ◽

Diagnostic Methods ◽

Virus Species ◽

Time Saving ◽

Robust Detection ◽

Highly Sensitive ◽

Reliable Differentiation ◽

World Organization ◽

Real Time Qpcr

Outbreaks of the three capripox virus species, namely lumpy skin disease virus, sheeppox virus, and goatpox virus, severely affect animal health and both national and international economies. Therefore, the World Organization for Animal Health (OIE) classified them as notifiable diseases. Until now, discrimination of capripox virus species was possible by using different conventional PCR protocols. However, more sophisticated probe-based real-time qPCR systems addressing this issue are, to our knowledge, still missing. In the present study, we developed several duplex qPCR assays consisting of different types of fluorescence-labelled probes that are highly sensitive and show a high analytical specificity. Finally, our assays were combined with already published diagnostic methods to a diagnostic workflow that enables time-saving, reliable, and robust detection, differentiation, and characterization of capripox virus isolates.

Download Full-text

A hybrid upconversion nanoprobe for ratiometric detection of aliphatic biogenic amines in aqueous medium

Nanoscale Advances ◽

10.1039/d0na00995d ◽

2021 ◽

Author(s):

Shilpi Jaiswal ◽

Subhankar Kundu ◽

Sujoy Bandyopadhyay ◽

Abhijit Patra

Keyword(s):

Aqueous Medium ◽

Biogenic Amines ◽

Real Time ◽

Inorganic Hybrid ◽

The Real ◽

Ratiometric Detection ◽

Real Time Detection

An organic–inorganic hybrid upconversion nanoprobe was developed for the real-time detection of aliphatic biogenic amines in an aqueous medium, adulterated milk, and rotten fish.

Download Full-text

Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2

Scientific Reports ◽

10.1038/s41598-021-88625-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Chukwunonso Onyilagha ◽

Henna Mistry ◽

Peter Marszal ◽

Mathieu Pinette ◽

Darwyn Kobasa ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Real Time ◽

Point Of Care ◽

Middle Eastern ◽

Turnaround Time ◽

Cross Reactivity ◽

Clinical Samples ◽

Diarrhea Virus ◽

Highly Sensitive ◽

Transmissible Gastroenteritis

AbstractThe coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.

Download Full-text

Highly sensitive real-time detection of intracellular oxidative stress and application in mycotoxin toxicity evaluation based on living single-cell electrochemical sensors

The Analyst ◽

10.1039/d0an02015j ◽

2021 ◽

Author(s):

Lu Gao ◽

Jiadi Sun ◽

Liping Wang ◽

Qigao Fan ◽

Gaowen Zhu ◽

...

Keyword(s):

Oxidative Stress ◽

Electrochemical Sensor ◽

Single Cell ◽

Real Time ◽

Electrochemical Sensors ◽

Living Cells ◽

Selective Detection ◽

Toxicity Evaluation ◽

Highly Sensitive ◽

Intracellular Oxidative Stress

Single-cell electrochemical sensor is used in the local selective detection of living cells because of its high spatial–temporal resolution and sensitivity, as well as its ability to obtain comprehensive cellular physiological states and processes.

Download Full-text

Real-Time In Vivo Bioluminescent Imaging for Evaluating the Efficacy of Antibiotics in a Rat Staphylococcus aureus Endocarditis Model

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.49.1.380-387.2005 ◽

2005 ◽

Vol 49 (1) ◽

pp. 380-387 ◽

Cited By ~ 68

Author(s):

Yan Q. Xiong ◽

Julie Willard ◽

Jagath L. Kadurugamuwa ◽

Jun Yu ◽

Kevin P. Francis ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Real Time ◽

Imaging System ◽

Bioluminescent Imaging ◽

Vancomycin Therapy ◽

Treatment Groups ◽

Highly Sensitive ◽

Relevant Animal Model ◽

And Control

ABSTRACT Therapeutic options for invasive Staphylococcus aureus infections have become limited due to rising antimicrobial resistance, making relevant animal model testing of new candidate agents more crucial than ever. In the present studies, a rat model of aortic infective endocarditis (IE) caused by a bioluminescently engineered, biofilm-positive S. aureus strain was used to evaluate real-time antibiotic efficacy directly. This strain was vancomycin and cefazolin susceptible but gentamicin resistant. Bioluminescence was detected and quantified daily in antibiotic-treated and control animals with IE, using a highly sensitive in vivo imaging system (IVIS). Persistent and increasing cardiac bioluminescent signals (BLS) were observed in untreated animals. Three days of vancomycin therapy caused significant reductions in both cardiac BLS (>10-fold versus control) and S. aureus densities in cardiac vegetations (P < 0.005 versus control). However, 3 days after discontinuation of vancomycin therapy, a greater than threefold increase in cardiac BLS was observed, indicating relapsing IE (which was confirmed by quantitative culture). Cefazolin resulted in modest decreases in cardiac BLS and bacterial densities. These microbiologic and cardiac BLS differences during therapy correlated with a longer time-above-MIC for vancomycin (>12 h) than for cefazolin (∼4 h). Gentamicin caused neither a reduction in cardiac S. aureus densities nor a reduction in BLS. There were significant correlations between cardiac BLS and S. aureus densities in vegetations in all treatment groups. These data suggest that bioluminescent imaging provides a substantial advance in the real-time monitoring of the efficacy of therapy of invasive S. aureus infections in live animals.

Download Full-text