Development of a New Method for Diagnosis of Rubella Virus Infection by Reverse Transcription-Loop-Mediated Isothermal Amplification

Mōri, Nobuo; Motegi, Yoshie; Shimamura, Yasushi; Ezaki, Takashi; Natsumeda, Tomo; Yonekawa, Toshihiro; Ota, Yoshinori; Notomi, Tsugunori; Nakayama, Tetsuo

doi:10.1128/jcm.00803-06

Cited by 26 publications

(15 citation statements)

References 27 publications

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“…These conditions are consistent with those found for other single-stranded RNA viruses such as the infectious hematopoietic necrosis virus (Gunimaladevi et al, 2005), the rift valley fever virus (Peyrefitte et al, 2008) and the Crimean Congo hemorrhagic fever virus (Osman et al, 2013). The results of the temperature optimization showed that Bst DNA polymerase effectively amplified the nucleic acid templates at temperatures of 63-66 • C, which is consistent with the classic reports on LAMP (Notomi et al, 2000;Mori et al, 2001Mori et al, , 2006. The effective amplification of FHMNV nucleic acid by RT-LAMP in a wide temperature range should greatly benefit the future application of the method under In lanes 1-8 the reaction was conducted using 10-fold serial dilutions of RNA from FHMNV infected fish of 1.6 × 10 5 to 0 fg, respectively.…”

Section: Discussionsupporting

confidence: 89%

Development and evaluation of reverse transcription loop-mediated isothermal amplification assay for the detection of the fathead minnow nidovirus

Zhang

Standish

Winters

et al. 2014

Journal of Virological Methods

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Fathead minnow nidovirus (FHMNV) is a serious baitfish-pathogenic virus in North America. Studies to trace the spread of the virus and determine its host range are hampered by the absence of reliable diagnostic assays. In this study, a one-step, reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed that targets a region in the FHMNV spike protein gene. The assay was optimized, and the best results were obtained at 8 mM of Mg(2+) with an incubation time of 40 min at 63 °C in the presence of calcein. The analytical sensitivity of the RT-LAMP method was estimated to be as low as 5 viral copies and was 1000-fold more sensitive than the conventional reverse transcription polymerase chain reaction (RT-PCR) method. The diagnostic sensitivity and specificity of the developed RT-LAMP assay versus the RT-PCR assay was 100% and 95.7%, respectively. A quantitative RT-LAMP of FHMNV with a high correlation coefficient (r(2)=0.9926) was also developed and the result of quantitation of viral copies in tissue samples of infected fish showed that the viral loads of the infected fish tissue samples reached up to 4.7×10(10) copies per mg. It is anticipated that the developed RT-LAMP and quantitative RT-LAMP methods will be instrumental for diagnosis and surveillance of FHMNV.

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Section: Discussionsupporting

confidence: 89%

Development and evaluation of reverse transcription loop-mediated isothermal amplification assay for the detection of the fathead minnow nidovirus

Zhang

Standish

Winters

et al. 2014

Journal of Virological Methods

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“…The only equipment needed for the LAMP reaction is a regular laboratory water bath or a heat block that furnishes a constant temperature of 62-65°C [16,17]. Thus, this method could potentially be a valuable tool for the rapid diagnosis of infectious diseases in both commercial and hospital laboratories of developing countries [6,7,11,15]. In this study, we developed the LAMP method for the detection of the Hepatitis C virus genome and compared the sensitivity of LAMP with those of nested-PCR using clinical samples.…”

Section: Introductionmentioning

confidence: 99%

Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hepatitis C virus

et al. 2012

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“…The loop-mediated isothermal amplification (LAMP) method has been shown to be highly accurate for the detection of DNA (4,20) and RNA (14,32,36) viruses, differentiation of viral serotypes and subtypes (17,21), and rapid diagnosis of bacterial infections (5). LAMP amplifies target nucleic acid under isother-mal conditions, usually between 60°C and 65°C (19).…”

mentioning

confidence: 99%

Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens

et al. 2009

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This paper reports on the development and validation of a real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) targeting the genomic large RNA segment of Rift Valley fever virus (RVFV). The set of six designed RT-LAMP primers identified strains of RVFV isolated in geographically distinct areas over a period of 50 years; there was no cross-reactivity with other genetically related and unrelated arboviruses. When testing serial sera and plasma from sheep experimentally infected with wild-type RVFV, there was 100% agreement between results of the RT-LAMP, a TaqMan-based real-time PCR, and virus isolation. Similarly, the assay had very high levels of diagnostic sensitivity and specificity when testing various clinical specimens from humans and animals naturally infected with the virus during recent outbreaks of the disease in Africa. The detection of specific viral genome targets in positive clinical specimens was achieved in less than 30 min. As a highly accurate, rapid, and very simple nucleic acid detection format, the RT-LAMP has the potential to be used in less-well-equipped laboratories in Africa and as a portable device during RVF outbreaks in remote areas, and it can be a valuable tool for the differential diagnosis of viral hemorrhagic fevers.

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Development of a New Method for Diagnosis of Rubella Virus Infection by Reverse Transcription-Loop-Mediated Isothermal Amplification

Cited by 26 publications

References 27 publications

Development and evaluation of reverse transcription loop-mediated isothermal amplification assay for the detection of the fathead minnow nidovirus

Development and evaluation of reverse transcription loop-mediated isothermal amplification assay for the detection of the fathead minnow nidovirus

Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hepatitis C virus

Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens

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