The Feline coronavirus (FCoV) is the etiological agent of feline infectious peritonitis (FIP), a lethal disease of felids. The role of molecular methods is controversial for the diagnosis of FIP, while essential for the identification of the shedders. Thus, a fast and inexpensive method for the detection of FCoV could be beneficial, especially in multicat environments. A reverse transcription loop mediated isothermal amplification (RT-LAMP) assay was developed. RNA extraction and RT-nPCR for FCoV were performed on thirty-two samples (11 faeces, 9 blood, 8 effusions, and 4 lymph nodes) collected from 27 cats. Six RT-LAMP primers were designed from the same conserved region of RT-nPCR, and the assay was run at 63°C for one hour. Results were evaluated through both agarose gel run and hydroxynapthol blue (HNB) dye and then compared with RT-nPCR results for the assessment of sensitivity and specificity. The overall specificity was 100%, but the sensitivity was 50% and 54.5% for agarose gel and HNB respectively. Therefore, RT-LAMP seems optimal to confirm the presence of the virus, but not applicable to exclude it.
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http://dx.doi.org/10.1016/j.jviromet.2017.01.009 | DOI Listing |
Mikrochim Acta
January 2025
Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China.
A point-of-care testing (POCT) assay based on commercial HCG strip was proposed for miRNA21 detection by integrating RCA-HCR cascaded isothermal amplification with CRISPR/Cas12a. Three modules were integrated in the proposed platform: target amplification module composed of rolling circle amplification (RCA) cascaded with hybridization chain reaction (HCR), signal transduction module composed of CRISPR/Cas12a combined with HCG-agarose gel beads probes, and signal readout module composed of commercial HCG strips. The proposed RCA-HCR-CRISPR/Cas12a-HCG strip assay for miRNA21 detection had high sensitivity, and the limit of detection was as low as 37 fM.
View Article and Find Full Text PDFNat Chem Biol
January 2025
Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, USA.
Cell-free systems are powerful synthetic biology technologies that can recapitulate gene expression and sensing without the complications of living cells. Cell-free systems can perform more advanced functions when genetic circuits are incorporated. Here we expand cell-free biosensing by engineering a highly specific isothermal amplification circuit called polymerase strand recycling (PSR), which leverages T7 RNA polymerase off-target transcription to recycle nucleic acid inputs within DNA strand displacement circuits.
View Article and Find Full Text PDFAnal Chem
January 2025
School of Chemistry and Chemical Engineering, Yancheng Institute of Technology, Yancheng 224051, P. R. China.
DNAzyme-based cascade networks are effective tools to achieve ultrasensitive detection of low-abundance miRNAs. However, their designs are complicated and costly, and the operation is time-consuming. Herein, a novel simple noncascade DNAzyme network is designed and its amplification effect is comparable to or even better than many cascading ones.
View Article and Find Full Text PDFVet Res Commun
January 2025
Faculty of Medical Technology, Prince of Songkla University, Songkhla, 90110, Thailand.
Staphylococcus pseudintermedius is a global animal pathogen. Traditional identification methods are time-consuming necessitating a more efficient approach. This study validated and enhanced the loop-mediated isothermal amplification (LAMP) technique by integration it with a lateral flow dipstick (LFD) assay for the detection of S.
View Article and Find Full Text PDFCureus
December 2024
Department of Respiratory Medicine, Fukuoka University Hospital, Fukuoka, JPN.
typically causes mild respiratory infections but can rarely lead to severe complications. We report a case of a 43-year-old immunocompetent male who presented with progressive dyspnea and respiratory failure with bilateral pulmonary infiltrates, refractory to outpatient treatment with azithromycin, ceftriaxone, and levofloxacin. Bronchoscopy revealed multiple white plaques in the trachea and diffuse alveolar hemorrhage.
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