LAMP real-time turbidity detection for fowl adenovirus

Abstract

Background Fowl adenovirus (FAdV) is an infectious agent that can cause jaundice, severe anaemia, dyspnoea and reproductive disorders in fowls. Since 2015, FAdV disease has been rapidly spreading among broiler chickens in China, where it has caused significant economic losses. In this study, a loop-mediated isothermal amplification (LAMP) real-time turbidity method with strong specificity to FAdV was established. Results The established assay was specific to FAdV-4, and the lower limit of detection was 75 copies/μL of extracted DNA. The positive detection rate for the suspected tissue samples was 33.3% (14/42), which was consistent with that of the real-time PCR. Conclusion The proposed LAMP method can quickly and accurately detect prevalent FAdV via real-time turbidity assay, thereby providing a diagnostic platform for the prevention and control of the FAdV disease.

A novel recombinase polymerase amplification assay for rapid detection of epidemic fowl adenovirus

“…Although these molecular diagnostics, comparing with traditional methods, has significantly improved in virology diagnosing accuracy, some disadvantages in respect of the operational complexity and thermocycler dependence are not well solved, limiting its use mostly in specialized and suitably equipped labs ( Ma et al, 2018 ). Newly developed methods, like LAMP, could detect FAdV isothermally with high sensitivity, but the temperature needed was very high (65 °C) and merely suitable for the specific strain (FAdV-4) ( Yuan et al, 2019 ; Zhai et al, 2019 ). While as the RPA assay putting into practice in Zika virus (ZIKV) and other virus detection, it enables rapid detection possible ( Babu et al, 2017 ; Vasileva Wand et al, 2018 ).…”

Toward a next-generation diagnostic tool: A review on emerging isothermal nucleic acid amplification techniques for the detection of SARS-CoV-2 and other infectious viruses

“…dNTPs are substrates of Bst polymerase enzymes, and their turnover leaves pyrophosphate anions in the system, which rapidly react with magnesium cations in the buffer [ 69 , 70 ]. This endpoint detection offers simple naked-eye monitoring of the LAMP products; however, commercial turbidimeters now allow quantitative detection in real-time [ 71 , 72 ].…”

Competitive SNP-LAMP probes for rapid and robust single-nucleotide polymorphism detection

“…Using the MT-CO2, ACTB, TP53 , and NOTCH1 LAMP primers listed in Table S2, we performed LAMP amplification from 10ng of MOLT-4 or SK-BR-3 total RNA in 25μL reactions for 90 minutes, as described above. We verified reaction completion by eye using turbidity 43 . Upon reaction completion, we added 10μg of RNase A (Thermo Fisher) and incubated at 37°C for 15 minutes to destroy cellular RNA.…”