Application of multiplex reverse transcription-ligase detection reaction-polymerase chain reaction (MRLP) mediated universal DNA microarray for detecting potato viruses in field samples

作者：Wang Ping; Guo Yao; Cheng Ju Hui; Dong Qin Fang; Ding Xian Feng; Guo Jiang Feng; Jiang Yong Hou^*

来源：European Journal of Plant Pathology, 2012, 132(2): 217-227.

DOI：10.1007/s10658-011-9865-4

• LDR-PCR
• Universal microarray
• Potato viruses
• Field samples
• Detection

To facilitate the comprehensive and unbiased analysis of viral prevalence in a given biological setting, we have applied conventional PCR and multiplex reverse transcription-ligase detection reaction-polymerase chain reaction (MRLP) based universal microarray for the identification of potato virus infections in potato samples from tubers and leaves. With this MRLP based microarray (MRLPM) technique, total RNAs of 79 field samples were reverse transcribed by random primers, then subjected to a ligase detection reaction (LDR) and asymmetric labelling PCR as templates, finally, the MRLP amplicons were analyzed by zip-code microarray hybridization and subsequent scanning. The concordance rates between the MRLPM and the PCR results were 92.50 to 98.73% for ten potato viruses. In addition, the positive incidence obtained by the MRLPM was higher than that by PCR for corresponding viruses in field samples. In conclusion, the method was able to simultaneously detect ten potato viruses from samples with and without symptoms. Thus, this assay can be used as a rapid and sensitive technique for routine potato virus detection in field surveillance and in plant quarantine.

• 出版日期2012-2
• 单位浙江理工大学