สืบค้นงานวิจัย
Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes
Arunrut N. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes
ผู้แต่ง / หัวหน้าโครงการ (EN): Arunrut N.
บทคัดย่อ (EN): Acute hepatopancreatic necrosis disease (AHPND) is a component cause of early mortality syndrome (EMS) of shrimp. In 2013, the causative agent was found to be unique isolates of Vibrio parahaemolyticus (VPAHPND) that contained a 69 kbp plasmid (pAP1) carrying binary Pir-like toxin genes PirvpA and PirvpB. In Thailand, AHPND was first recognized in 2012, prior to knowledge of the causative agent, and it subsequently led to a precipitous drop in shrimp production. After VPAHPND was characterized, a major focus of the AHPND control strategy was to monitor broodstock shrimp and post larvae for freedom from VPAHPND by nucleic acid amplification methods, most of which required use of expensive and sophisticated equipment not readily available in a shrimp farm setting. Here, we describe a simpler but equally sensitive approach for detection of VPAHPND based on loop-mediated isothermal amplification (LAMP) combined with unaided visual reading of positive amplification products using a DNA-functionalized, ssDNA-labled nanogold probe (AuNP). The target for the special set of six LAMP primers used was the VPAHPND PirvpA gene. The LAMP reaction was carried out at 65°C for 45 min followed by addition of the red AuNP solution and further incubation at 65°C for 5 min, allowing any PirvpA gene amplicons present to hybridize with the probe. Hybridization protected the AuNP against aggregation, so that the solution color remained red upon subsequent salt addition (positive test result) while unprotected AuNP aggregated and underwent a color change from red to blue and eventually precipitated (negative result). The total assay time was approximately 50 min. The detection limit (100 CFU) was comparable to that of other commonly-used methods for nested PCR detection of VPAHPND and 100-times more sensitive than 1-step PCR detection methods (104 CFU) that used amplicon detection by electrophoresis or spectrophotometry. There was no cross reaction with DNA templates derived from non-AHPND bacteria commonly found in shrimp ponds (including other Vibrio species). The new method significantly reduced the time, difficulty and cost for molecular detection of VPAHPND in shrimp hatchery and farm settings. © 2016 Arunrut et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-84962440521&doi=10.1371%2fjournal.pone.0151769&partnerID=40&md5=4c16b98410079f80c2925cb8e1433d32
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): Vibrio parahaemolyticus
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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Arunrut N.. "Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes". ไม่ระบุหน่วยงาน. ม.ป.ป.

Arunrut N.. "Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNAFunctionalized gold nanoparticles as probes
Arunrut N.
มหาวิทยาลัยมหิดล
ไม่ระบุวันที่เผยแพร่
In situ DIG-labeling, loop-mediated DNA Amplification (ISDL) for highly sensitive detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) การพัฒนาวิธีการตรวจโรคเมลิออยโดสิสด้วย loop-mediated isothermal amplification (LAMP) Development of loop-mediated isothermal amplification (LAMP) for diagnosing melioidosis Double-Loop-Mediated Isothermal Amplification (D-LAMP) using colourimetric gold nanoparticle probe for rapid detection of infectious Penaeus stylirostris densovirus (PstDNV) with reduced false-positiv Development of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) for the detection of the causative agent of covid-19 In situ SERS detection of emulsifiers at lipid interfaces using label-free amphiphilic gold nanoparticles การพัฒนาวิธีการตรวจโรคเมลิออยโดสิสและเลปโตสไปโรสิสด้วย loop-mediated isothermal amplification (LAMP) Development of loop-mediated isothermal amplification (LAMP) for diagnosing melioidosis and leptosp Comparison of Loop-Mediated Isothermal Amplification, Microscopy, Culture, and PCR for Diagnosis of Pulmonary Tuberculosis The performance of in-house loop-mediated isothermal amplification for rapid detection of Mycobacterium tuberculosis in sputum sample comparing to Xpert MTB/RIF, microscopy and culture Development of the Rapid Test Kit for the Identification of Campylobacter spp. Based on Loop-mediated Isothermal Amplification (LAMP) in Combination with a Lateral Flow Dipstick (LFD) and Gold Nano-DN Detection of Campylobacter DNA using magnetic nanoparticles coupled with PCR and a colorimetric end-point system
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