Application of high resolution melt (HRM) analysis for duplex detection of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) in shrimp

Senapin, Saengchan; Molthathong, Sudkhate; Phiwsaiya, Kornsunee; Jaengsanong, Chatlada; Chuchird, Niti

doi:10.1016/j.mcp.2010.06.003

Cited by 23 publications

(6 citation statements)

References 31 publications

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“…In this paper, we suggest a real‐time polymerase chain reaction high resolution melt (PCR HRM) assay to detect trace biological material from P. cubensis as an inexpensive alternative for situations where collected samples are too scarce for conventional analytical drug testing methods. The PCR HRM method has been demonstrated for marijuana and “legal high” drugs among other species differentiation and identification assays including a preliminary melt assay for Psilocybe with a melt temperature of 82°C presented at a recent AAFS meeting, although the primer sequences, developmental validation, and casework testing were not discussed . We also demonstrate a PCR HRM duplex assay that can simultaneously detect marijuana and P. cubensis DNA.…”

mentioning

confidence: 99%

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Cowan

Elkins

2017

Journal of Forensic Sciences

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Psilocybe cubensis, or "magic mushroom," is the most common species of fungus with psychedelic characteristics. Two primer sets were designed to target Psilocybe DNA using web-based software and NBCI gene sequences. DNA was extracted from eighteen samples, including twelve mushroom species, using the Qiagen DNeasy Plant Mini Kit. The DNA was amplified by the polymerase chain reaction (PCR) using the primers and a master mix containing either a SYBR Green I, Radiant™ Green, or LCGreen Plus intercalating dye; amplicon size was determined using agarose gel electrophoresis. The PCR assays were tested for amplifiability, specificity, reproducibility, robustness, sensitivity, and multiplexing with primers that target marijuana. The observed high resolution melt (HRM) temperatures for primer sets 1 and 7 were 78.85 ± 0.31°C and 73.22 ± 0.61°C, respectively, using SYBR Green I dye and 81.67 ± 0.06°C and 76.04 ± 0.11°C, respectively, using Radiant™ Green dye.

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confidence: 99%

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Cowan

Elkins

2017

Journal of Forensic Sciences

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“…Unlike endpoint PCR plus electrophoresis, real‐time PCR HRM assays are relatively quick (<20 min for HRM) and assay cost is affordable (approximately one USD per sample). PCR HRM assays have differentiated species including 10 ungulate species , Brucella species , two bollworm species , two shrimp viruses , bacterial community compositions , and three bacterial food‐borne pathogens . PCR HRM has also been used to differentiate body fluids and to differentiate STRs .…”

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confidence: 99%

“…Second, the method is cheaper than previous assays , as it requires no expensive dye‐labeled primers. Third, the assays are capable of detecting the presence of four plant species simultaneously using HRM peak differentiation by melt temperature (previously two and three species were simultaneously identified using this method). Fourth, differentiation by melt temperature is straightforward and intuitive, unlike HRM difference plots or traces of melt curve kinetics .…”

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confidence: 99%

Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay^,

Elkins

Perez

Quinn

2016

Journal of Forensic Sciences

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The international prevalence of "legal high" drugs necessitates the development of a method for their detection and identification. Herein, we describe the development and validation of a tetraplex multiplex real-time polymerase chain reaction (PCR) assay used to simultaneously identify morning glory, jimson weed, Hawaiian woodrose, and marijuana detected by high-resolution melt using LCGreen Plus . The PCR assay was evaluated based on the following: (i) specificity and selectivity-primers were tested on DNA extracted from 30 species and simulated forensic samples, (ii) sensitivity-serial dilutions of the target DNA were prepared, and (iii) reproducibility and reliability-sample replicates were tested and remelted on different days. The assay is ideal for cases in which inexpensive assays are needed to quickly detect and identify trace biological material present on drug paraphernalia that is too compromised for botanical microscopic identification and for which analysts are unfamiliar with the morphology of the emerging "legal high" species.

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“…Nucleic acid concentration and quality were measured by spectrophotometric analysis at 260 and 280 nm. Viral infection was confirmed by respective IQ2000 Detection kits (GeneReach Corp., Taiwan) except for IMNV and MrNV that were detected according to previously described RT-PCR protocols [17,18].…”

Section: Viral Infected Shrimp Samples and Nucleic Acid Extractionmentioning

confidence: 99%

Shrimp laminin receptor binds with capsid proteins of two additional shrimp RNA viruses YHV and IMNV

Busayarat

Senapin

Tonganunt

et al. 2011

Fish & Shellfish Immunology

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Application of high resolution melt (HRM) analysis for duplex detection of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) in shrimp

Cited by 23 publications

References 31 publications

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay^,

Shrimp laminin receptor binds with capsid proteins of two additional shrimp RNA viruses YHV and IMNV

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Application of high resolution melt (HRM) analysis for duplex detection of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) in shrimp

Cited by 23 publications

References 31 publications

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Detection and Identification of Psilocybe cubensis DNA Using a Real‐Time Polymerase Chain Reaction High Resolution Melt Assay,

Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay,

Shrimp laminin receptor binds with capsid proteins of two additional shrimp RNA viruses YHV and IMNV

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Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay^,