Specific detection of reverse transcription-loop-mediated isothermal amplification amplicons for Taura syndrome virus by colorimetric dot–blot hybridization

Teng, P. K.; Chen, Chu-Liang; Sung, Ping-Feng; Lee, Fu-Chun; Ou, Bor-Rung; Lee, Pei-Yu

doi:10.1016/j.jviromet.2007.07.027

Cited by 55 publications

(32 citation statements)

References 35 publications

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“…This phenomenon has been widely reported in the literature 36-39 , and is usually circumvented by optimization of assay conditions and setting an assay cutoff time far before these events are likely to occur. Nonetheless, because our ultimate assay detection method is based on primer-tagged probes, it was important to ensure that a positive result on the LFD strip correlated to a LAMP product specific to our target sequence.…”

Section: Resultsmentioning

confidence: 99%

A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples

et al. 2016

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Paper diagnostics have successfully been employed to detect the presence of antigens or small molecules in clinical samples through immunoassays; however, the detection of many disease targets relies on the much higher sensitivity and specificity achieved via nucleic acid amplification tests (NAAT). The steps involved in NAAT have recently begun to be explored in paper matrices, and our group, among others, has reported on paper-based extraction, amplification, and detection of DNA and RNA targets. Here, we integrate these paper-based NAAT steps onto a single paperfluidic chip in a modular, foldable system that allows for fully integrated fluidic handling from sample to result. We showcase the functionality of the chip by combining nucleic acid isolation, isothermal amplification, and lateral flow detection of human papillomavirus (HPV) 16 DNA directly from crude cervical specimens in under 1 hour for rapid, early detection of cervical cancer. The chip is made entirely of paper and adhesive sheets, making it low-cost, portable, and disposable, and offering the potential for a point-of-care molecular diagnostic platform even in remote and resource-limited settings.

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Section: Resultsmentioning

confidence: 99%

A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples

et al. 2016

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“…However, it is in general more challenging to optimize and multiplex LAMP assays because they require four to six primers to hybridize with six to eight areas in one target area. Detection of target amplicon-specific signals would reduce greatly the risks of false positive results derived from non-specific LAMP amplicons [23], [26], [30], [31]. However, to our best knowledge, portable devices for target amplicon-specific detection are not available currently for field application of LAMP assays.…”

Section: Discussionmentioning

confidence: 99%

Validation of a Commercial Insulated Isothermal PCR-based POCKIT Test for Rapid and Easy Detection of White Spot Syndrome Virus Infection in Litopenaeus vannamei

Tsai

Wang

et al. 2014

PLoS ONE

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Timely pond-side detection of white spot syndrome virus (WSSV) plays a critical role in the implementation of bio-security measures to help minimize economic losses caused by white spot syndrome disease, an important threat to shrimp aquaculture industry worldwide. A portable device, namely POCKIT™, became available recently to complete fluorescent probe-based insulated isothermal PCR (iiPCR), and automatic data detection and interpretation within one hour. Taking advantage of this platform, the IQ Plus™ WSSV Kit with POCKIT system was established to allow simple and easy WSSV detection for on-site users. The assay was first evaluated for its analytical sensitivity and specificity performance. The 95% limit of detection (LOD) of the assay was 17 copies of WSSV genomic DNA per reaction (95% confidence interval [CI], 13 to 24 copies per reaction). The established assay has detection sensitivity similar to that of OIE-registered IQ2000™ WSSV Detection and Protection System with serial dilutions of WSSV-positive Litopenaeus vannamei DNA. No cross-reaction signals were generated from infectious hypodermal and haematopoietic necrosis virus (IHHNV), monodon baculovirus (MBV), and hepatopancreatic parvovirus (HPV) positive samples. Accuracy analysis using700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5% (95% CI: 90.61–95.56%) and specificity 97% (95% CI: 94.31–98.50%). Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.

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“…Specificity and efficiency was found be highly dependent on Mg 2+ concentration. Mg 2+ is known to affect primer annealing (Nie 2005;Gunimaladevi et al 2005;Teng et al 2007;Liu et al 2010) and it is possible that the LAMP conditions make those conditions too stringent for primers to anneal to the slight variations in sequence. RT-LAMP ideally should be able to detect all strains of the GLRaV-3 and as such it is important that the conditions not be too stringent.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of Grapevine leafroll-associated virus type 3 using a reverse transcription loop-mediated amplification method

Walsh

Pietersen

2013

Journal of Virological Methods

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Specific detection of reverse transcription-loop-mediated isothermal amplification amplicons for Taura syndrome virus by colorimetric dot–blot hybridization

Cited by 55 publications

References 35 publications

A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples

A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples

Validation of a Commercial Insulated Isothermal PCR-based POCKIT Test for Rapid and Easy Detection of White Spot Syndrome Virus Infection in Litopenaeus vannamei

Rapid detection of Grapevine leafroll-associated virus type 3 using a reverse transcription loop-mediated amplification method

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