Reverse Transcription Loop-Mediated Isothermal Amplification of DNA for Detection of Potato virus Y

Plant Disease
Xianzhou Nie

Abstract

A reverse transcription loop-mediated isothermal amplification of DNA (RT-LAMP) for detection of Potato virus Y (PVY) was developed. In this procedure, a set of four primers matching a total of six sequences of the coat protein (CP) gene of PVY were designed in such a way that a loop could be formed and elongated during DNA amplification. Using PVY CP complementary DNA clones as templates, the LAMP reaction was optimized by adjusting the concentrations of MgSO4, dNTPs, and Bst DNA polymerase. The effects of fragment length of target DNA on LAMP also were investigated. Two-step and one-step RT-LAMPs were performed using RNA extracts of various PVY cultures, and the results were correlated with two-step reverse transcription polymerase chain reaction (RT-PCR) for detection of PVY. Further, the turbidity caused by precipitation of magnesium pyrophosphate formed in positive RT-LAMP reactions was used to measure the amplification by utilizing a time-saving spectrophotometric method. The one-step RT-LAMP-turbidity method gave results comparable with the two-step RT-PCR method for detection of PVY from potato leaf and tuber samples. Of the total 240 samples, 234 were diagnosed similarly by both methods.

References

Jun 28, 2000·Nucleic Acids Research·T NotomiT Hase
Nov 16, 2001·Biochemical and Biophysical Research Communications·Y MoriT Notomi
Jul 30, 2002·Molecular and Cellular Probes·K NagamineT Notomi
Sep 25, 2003·Archives of Virology·S FukutaY Ishimoto
Jan 13, 2004·Journal of Clinical Microbiology·Manmohan ParidaKouichi Morita
Jan 1, 1993·Annual Review of Phytopathology·J M Henson, R French

❮ Previous
Next ❯

Citations

Jan 19, 2018·Acta Histochemica Et Cytochemica·Ibu MatsuzakiShin-Ichi Murata
Jan 1, 2006·European Journal of Plant Pathology·Rick MumfordIan Barker
Mar 8, 2020·International Journal of Molecular Sciences·Sumyya WaliullahMd Emran Ali
Dec 16, 2020·International Journal of Molecular Sciences·Nikolaos NikoloudakisGeorgios Tsaniklidis
Jul 7, 2010·Journal of Virological Methods·Cheng-Ping KuanHung-Chang Huang
Nov 2, 2021·World Journal of Microbiology & Biotechnology·Chuan WuYang He

❮ Previous
Next ❯

Related Concepts

Related Feeds

Birth Defects

Birth defects encompass structural and functional alterations that occur during embryonic or fetal development and are present since birth. The cause may be genetic, environmental or unknown and can result in physical and/or mental impairment. Here is the latest research on birth defects.