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Demonstration of a very inexpensive, turbidimetric, real-time, RT-LAMP detection platform using shrimp laem-singh virus (LSNV) as a model
Arunrut N. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Demonstration of a very inexpensive, turbidimetric, real-time, RT-LAMP detection platform using shrimp laem-singh virus (LSNV) as a model
ผู้แต่ง / หัวหน้าโครงการ (EN): Arunrut N.
บทคัดย่อ (EN): Rapid and accurate detection of pathogens under field laboratory conditions is necessary for effective control of veterinary pathogens. Here we describe a prototype, portable, pathogen detection device developed for single tube, real-time, reverse transcription, loop-mediated isothermal amplification (RT-LAMP) using Laem-Singh virus (LSNV) as a model. LSNV is an RNA virus and a component cause of growth retardation in black tiger shrimp. We chose its RNA-dependent RNA polymerase (RdRp) gene as the target for our tests. The basis for detection was measurement of turbidity arising from formation of a white, insoluble magnesium pyrophosphate precipitate byproduct upon amplification of the RdRp target sequence from 100 ng template RNA extracted from shrimp. The measurement device consisted of a heating block to maintain constant temperature in the RT-LAMP reaction for 8 Eppindorf sample tubes, a light-emitting diode (LED) light source providing red light emission at 650 nm wavelength to pass through sample tubes, a light dependent resistance (LDR) photo-detector and a software program to report turbidity events and could potentially be marketed for under US$3000. The device was connected to a computer to display real-time results in a variety of formats. The optimized protocol for LSNV detection consisted of incubation of the sample tubes at 65°C for 1 h during which turbidity was continuously measured, and quantitative results could be obtained by reaction time measurement. The sensitivity of detection was comparable to that of conventional nested RT-PCR and there was no cross reaction with other common shrimp viruses. The device was used for quantitative measurement of relative copy numbers of LSNV RdRp in 8 shrimp tissues and they were found to be highest in the gills followed in order by the lymphoid organ and hemolymph (p<0.05). This platform can be easily adapted for detection of other pathogens under field laboratory settings. © 2014 Arunrut et al.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-84907450548&doi=10.1371%2fjournal.pone.0108047&partnerID=40&md5=5d90d66927fb5e1df73d246164a37298
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): RNA Viruses
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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Arunrut N.. "Demonstration of a very inexpensive, turbidimetric, real-time, RT-LAMP detection platform using shrimp laem-singh virus (LSNV) as a model". ไม่ระบุหน่วยงาน. ม.ป.ป.

Arunrut N.. "Demonstration of a very inexpensive, turbidimetric, real-time, RT-LAMP detection platform using shrimp laem-singh virus (LSNV) as a model". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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Demonstration of a very inexpensive, turbidimetric, real-time, RT-LAMP detection platform using shrimp laem-singh virus (LSNV) as a model
Arunrut N.
มหาวิทยาลัยมหิดล
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