000			02376nab a22003137a 4500
001			G95217
003			MX-TxCIM
005			20171220113530.0
008			121211b |||p||p||||||| |z||| |
022			_a1448-6032 (Revista en electrónico)
022	0		_a0815-3191
024	8		_ahttps://doi.org/10.1071/AP08088
040			_aMX-TxCIM
041	0		_aEn
100	1		_aMui-Keng Tan
245	0	0	_aA one-tube fluorescent assay for the quarantine detection and identification of Tilletia indica and other grass bunts in wheat
260			_c2009
500			_aPeer-review: Yes - Open Access: Yes|http://science.thomsonreuters.com/cgi-bin/jrnlst/jlresults.cgi?PC=MASTER&ISSN=0815-3191
520			_aA molecular assay with enhanced specificity and sensitivity has been developed to assist in the surveillance of Karnal bunt, a quarantineable disease with a significant impact on international trade. The protocol involves the release of DNA from spores, PCR amplification to enrich Tilletia-specific templates from released DNA and a five-plex, real-time PCR assay to detect, identify and distinguish T. indica and other Tilletia species (T. walkeri, T. ehrhartae, T. horrida and a group comprising T. caries, T. laevis, T. contraversa, T. bromi and T. fusca) in wheat grains. This fluorescent molecular tool has a detection sensitivity of one spore and thus bypasses the germination step, which in the current protocol is required for confirmation when only a few spores have been found in grain samples. The assay contains five dual-labelled, species-specific probes and associated species-specific primer pairs in a PCR mix in one tube. The different amplification products are detected simultaneously by five different fluorescence spectra. This specific and sensitive assay with reduced labour and reagent requirements makes it an effective and economically sustainable tool to be used in a Karnal bunt surveillance program. This protocol will also be valuable for the identification of some contaminant Tilletia sp. in wheat grains.
546			_aEnglish
700	1		_aGhalayini, A., _ecoaut.
700	1		_aJianping Yi, _ecoaut.
700	1		_aPriest, M., _ecoaut.
700	1		_aSharma, I., _ecoaut. _91884
700	1		_aShivas, R., _ecoaut.
700	1		_aWright, D., _ecoaut.
773	0		_tAustralasian Plant Pathology _gv. 38, no. 2, p. 101-109
942			_cJA _2ddc
999			_c28494 _d28494