tetano
Editor, Senior Moderator
Bing Du Xue Bao. 2013 Sep;29(5):488-94.
[Establish and optimization of real-time fluorescent reverse transcription loop-mediated isothermal amplification for detection of avian influenza H5 hemagglutinin gene].
[Article in Chinese]
Liu Y1, Tang JM2, Tao H2, Sun J3, Lu TK2, Liao LS2, Liu JL2, Zeng SL2, Cao CF2, Zhang CH2, Ruan ZX2, Lv JQ2, Yang JX2, Hua QY2, Chen ZL4, Qin ZF2.
Author information
Abstract
H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye. The results showed the sensitivity of this method was 100 times higher than that of standard real-time fluorescent RT-PCR, and the detection limit was one copy of the gene per reaction. This method had no cross-reactivity with other common avian respiratory tract infectious disease-related pathogens such as IBV and NDV. The present study suggested Calcein was the optimum dye. Small-scale tests suggested this method was reliable for survey monitoring of AIV-H5 on the spot, indicating its potential applications in field investigation.
PMID:
24386836
[PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/24386836
[Establish and optimization of real-time fluorescent reverse transcription loop-mediated isothermal amplification for detection of avian influenza H5 hemagglutinin gene].
[Article in Chinese]
Liu Y1, Tang JM2, Tao H2, Sun J3, Lu TK2, Liao LS2, Liu JL2, Zeng SL2, Cao CF2, Zhang CH2, Ruan ZX2, Lv JQ2, Yang JX2, Hua QY2, Chen ZL4, Qin ZF2.
Author information
Abstract
H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye. The results showed the sensitivity of this method was 100 times higher than that of standard real-time fluorescent RT-PCR, and the detection limit was one copy of the gene per reaction. This method had no cross-reactivity with other common avian respiratory tract infectious disease-related pathogens such as IBV and NDV. The present study suggested Calcein was the optimum dye. Small-scale tests suggested this method was reliable for survey monitoring of AIV-H5 on the spot, indicating its potential applications in field investigation.
PMID:
24386836
[PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/24386836