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040			_cCICY
090			_aB-11861
245	1	0	_aLeishmania parasite detection and quantification sing PCR-ELISA
490	0		_vNature Protocols, 5, p.1074-1080, 2010
520	3		_aThis protocol describes an improved and optimized PCR -EL ISA method for detection and quantification of Leishmania parasites in host tissues. Unlike other DNA -based assays, this method uses digoxigenin- and biotin-labeled primers. This eliminates the need for a separate step of hybridization of the PCR product with labeled probes. The PCR product is detected using sandwich EL ISA with antidigoxigenin-detecting antibodies. Primers are complementary to the kinetoplast minicircle conserved region of parasite DNA , allowing the detection of several Leishmania species. For measurement of a wide range of parasite concentrations, ±25 cycles were optimal. The sensitivity of this technique is 0.3 fg of parasite DNA per reaction in 40-cycle PCR -EL ISA , corresponding to 0.004 parasites. DNA preparation by a standard TR I reagent procedure takes about 4 h. When DNA is prepared, a single person can test a large number of samples (at least 150)in a maximum of 7 h. This method might also be suitable for detecting and quantifying other pathogens, especially for detecting small di fferences in pathogen numbers.
700	1	2	_aKobets, T.
700	1	2	_aBadalová, J.
700	1	2	_aGrekov, I.
700	1	2	_aHavelková, H.
700	1	2	_aSvobodová, M.
700	1	2	_aLipoldová, M.
856	4	0	_uhttps://drive.google.com/file/d/1xx67URoB50xAcQ1Z_4jmuF_JRuv_pHya/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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