2015
DOI: 10.1016/j.jviromet.2014.10.020
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Advanced loop-mediated isothermal amplification method for sensitive and specific detection of Tomato chlorosis virus using a uracil DNA glycosylase to control carry-over contamination

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Cited by 63 publications
(38 citation statements)
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“…Therefore, precautions should be taken such as careful opening and closing of reaction tubes, frequent changing the gloves, and separating the progress of pre- and post-RPA amplification. Furthermore, replacement of dTTP with dUTP may help prevent such carryover contamination as demonstrated in PCR and LAMP assays [35, 36]. Our results showed that the developed RPA assay could tolerate base mismatch in some degree as the primes could not completely match with the two PCV2 strains used in our work based on sequence alignment, and it may detect PCV-2 strains in some degree of variability.…”
Section: Discussionmentioning
confidence: 81%
“…Therefore, precautions should be taken such as careful opening and closing of reaction tubes, frequent changing the gloves, and separating the progress of pre- and post-RPA amplification. Furthermore, replacement of dTTP with dUTP may help prevent such carryover contamination as demonstrated in PCR and LAMP assays [35, 36]. Our results showed that the developed RPA assay could tolerate base mismatch in some degree as the primes could not completely match with the two PCV2 strains used in our work based on sequence alignment, and it may detect PCV-2 strains in some degree of variability.…”
Section: Discussionmentioning
confidence: 81%
“…Therefore, they are not ideal approaches for controlling carryover contamination in LAMP reaction33. Compared with the aforementioned methods, the UDG-mediated digestion of dUTP-incorporated contaminant DNA is simple, inexpensive, and does not require substantial modification of existing protocols42.…”
Section: Discussionmentioning
confidence: 99%
“…This decreases the Mg 2+ ion concentration, which is indicated by a colour change from violet to blue in the presence of HNB (Goto et al 2009). LAMP detection assays have been developed for several pathosystems, including oomycetes, fungi, bacteria and viruses (Harper et al 2010;Dai et al 2012;B€ uhlmann et al 2013;Chen et al 2013;Moradi et al 2013;Duan et al 2014;Kil et al 2015;Thiessen et al 2015). A LAMP assay for the detection of P. infestans, an organism which significantly impacts potato and tomato production globally, has not yet been developed.…”
Section: Introductionmentioning
confidence: 99%