کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
867837 909793 2011 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Development of multiplex reverse transcription-ligase detection reaction-polymerase chain reaction (MRLP) mediated universal DNA microarray for diagnostic platform
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Development of multiplex reverse transcription-ligase detection reaction-polymerase chain reaction (MRLP) mediated universal DNA microarray for diagnostic platform
چکیده انگلیسی

A multiplex reversal transcription-ligase detection reaction-polymerase chain reaction (MRLP) based universal microarray for the simultaneous pathogens detection was established by using potato viruses as a model. The proposed procedure integrated LDR for multiplicity and specificity, PCR amplification by universal primers for sensitivity, which required design of upstream and downstream LDR probes specific for each virus, and subsequent Zip-code microarray for multiplex and specific identification. Each MRLP fragments carried a unique sequence (complementary Zip-code sequence, cZip-code) which identified a virus by addressed to the location on the microarray where the Zip-code sequence has been spotted. Such Zip-code microarray and universal primers are therefore “universal” being unrelated to a specific molecular analyte. With this technique, target RNAs of ten potato viruses were reversal transcribed by random primer in a single reaction, then subjected to LDR and asymmetric labeling PCR as template, finally, the MRLP amplicons were analyzed by microarray hybridization and subsequent scanning. The technique platform was optimized and evaluated by using reference samples and artificial samples, which can specifically detect down to 3 copies of single or mixed plasmid templates. Due to its universality, multiplexing, specificity and sensitivity, this method can be recommended for simultaneously detecting a large number of different target types in related fields.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 26, Issue 8, 15 April 2011, Pages 3719–3724
نویسندگان
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