共查询到20条相似文献,搜索用时 420 毫秒
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Sappat A Jaroenram W Puthawibool T Lomas T Tuantranont A Kiatpathomchai W 《Journal of virological methods》2011,175(2):141-148
In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg2P2O7). The device incorporated a heating block that maintained an optimal temperature of 63 °C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10 min for rapid RNA preparation with 30 min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1 h compared to 4-8 h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field. 相似文献
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Development of reverse-transcription loop-mediated isothermal amplification for the detection of infectious bursal disease virus 总被引:1,自引:0,他引:1
Jiangtao Xu Zhenmei Zhang Yanbo Yin Shangjin Cui Shouzhen Xu Yanyan Guo Jida Li Jianlin Wang Xingcai Liu Limin Han 《Journal of virological methods》2009,162(1-2):267-271
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Loop-mediated isothermal amplification (LAMP) of DNA is a novel technique for the amplification of DNA under isothermal conditions. For the first time, we applied this method to develop a simple and sensitive tool for the detection of Pepino mosaic virus (PepMV). PepMV is an emerging pathogen that causes yield and quality losses in tomato crops. Specific RT-LAMP primers for PepMV detection were designed based on triple gene block sequences. The reaction was performed in a single tube at 65 °C for 30 min. The RT-LAMP assay is easy to perform and inexpensive, and it may be applied in the rapid and specific diagnosis of PepMV. 相似文献
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Arunrut N Prombun P Saksmerprome V Flegel TW Kiatpathomchai W 《Journal of virological methods》2011,171(1):21-25
Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is an important shrimp pathogen that causes mortality in Penaeus stylirostris and stunting (called runt deformity syndrome or RDS) in Penaeus vannamei. Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acids under isothermal conditions. It can be combined with a chromatographic lateral flow dipstick (LFD) for highly specific, rapid and simple visual detection of IHHNV-specific amplicons. Using this protocol, a 30-min amplification followed by 5 min hybridization with an FITC-labeled DNA probe and 5 min LFD resulted in visualization of DNA amplicons trapped at the LFD test line. Thus, 10 min for rapid DNA extraction followed by LAMP combined with LFD detection resulted in a total assay time of approximately 50 min. Detection sensitivity was comparable to other methods used commonly for nested PCR detection of IHHNV but had the additional advantages of reduced assay time, confirmation of amplicon identity by hybridization and elimination of electrophoresis with carcinogenic ethidium bromide. 相似文献
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Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus 总被引:11,自引:0,他引:11 下载免费PDF全文
Parida MM Santhosh SR Dash PK Tripathi NK Saxena P Ambuj S Sahni AK Lakshmana Rao PV Morita K 《Journal of clinical microbiology》2006,44(11):4172-4178
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Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus 总被引:24,自引:0,他引:24 下载免费PDF全文
Hong TC Mai QL Cuong DV Parida M Minekawa H Notomi T Hasebe F Morita K 《Journal of clinical microbiology》2004,42(5):1956-1961
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Early Detection of Dengue Virus by Use of Reverse Transcription-Recombinase Polymerase Amplification
Boon-Teong Teoh Sing-Sin Sam Kim-Kee Tan Mohammed Bashar Danlami Meng-Hooi Shu Jefree Johari Poh-Sim Hooi David Brooks Olaf Piepenburg Oliver Nentwich Annelies Wilder-Smith Leticia Franco Antonio Tenorio Sazaly AbuBakar 《Journal of clinical microbiology》2015,53(3):830-837
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Alvin Jin-Cherng Eun Liqun Huang Fook Tim Chew Sam Fong-Yau Li Sek Man Wong 《Journal of virological methods》2002,99(1-2):71-79
Quartz crystal microbalance (QCM) immunosensors are based on the principle that adsorption of substances on the surface of a quartz crystal changes its resonance oscillation frequency. A QCM immunosensor was developed for the detection of both cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) by pre-coating the QCMs with virus-specific antibodies. Upon binding of virions in either purified form or crude sap of infected orchids with the immobilised virus antibodies, the increase in mass at the QCM surface resulted in a reduction in the frequency of resonance oscillation in a manner dependent upon the amount of virus bound. The QCM was able to detect as low as 1 ng each of the two orchid viruses. This detection sensitivity is comparable to enzyme linked immunosorbent assay (ELISA) but the assay is faster. This immunoassay was shown to be specific, sensitive, rapid and economical, thus providing a viable alternative to virus detection methods. This is the first report using QCM immunosensors to detect plant viruses. 相似文献
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Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loop-mediated isothermal amplification assay 总被引:17,自引:0,他引:17
Parida M Horioke K Ishida H Dash PK Saxena P Jana AM Islam MA Inoue S Hosaka N Morita K 《Journal of clinical microbiology》2005,43(6):2895-2903
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