Visual detection of IS6110 of Mycobacterium tuberculosis in sputum samples using a test based on colloidal gold and latex beads P. Upadhyay, M. Hanif,

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Presentation transcript:

Visual detection of IS6110 of Mycobacterium tuberculosis in sputum samples using a test based on colloidal gold and latex beads P. Upadhyay, M. Hanif, S. Bhaskar Clinical Microbiology and Infection Volume 12, Issue 11, Pages 1118-1122 (November 2006) DOI: 10.1111/j.1469-0691.2006.01515.x Copyright © 2006 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 1 A, the sequences of the target strand, the nucleotide probe labelled with latex beads, and the nucleotide probe labelled with gold colloid. B, after a single mutation in the target strand, the nucleotide labelled with latex beads does not bind. Numbers indicate the position of the sequences in the IS6110 target sequence. Clinical Microbiology and Infection 2006 12, 1118-1122DOI: (10.1111/j.1469-0691.2006.01515.x) Copyright © 2006 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 2 Examples of results obtained with the gold colloid–latex bead test and PCR. A, photographs of filters showing the spots obtained with blank, 25 pg, 250 pg and 2.5 ng synthetic target nucleotide after washing (top row) and silver enhancement (bottom row). The spots are almost invisible after washing but become visible after silver enhancement. B, comparison of results obtained with PCR and the gold colloid–latex bead test showing the amplification obtained with 5 ng, 50 ng, 0.5 µg, 5 µg and 50 µg of target DNA (lanes 2, 3, 4, 5 and 7, respectively; lane 1, negative control; lane 6, 100-bp DNA ladder). Two-µL of amplified product was loaded in each lane. Spots obtained with the same amount of DNA are shown below every lane. C, variation in the intensity of these spots, showing an average of five spots and the standard deviations. D, comparison of the intensity of spots obtained from different types of DNA mixtures. The sensitivity of the test remained unchanged in a mixture of similar DNA or large amounts of random DNA. Clinical Microbiology and Infection 2006 12, 1118-1122DOI: (10.1111/j.1469-0691.2006.01515.x) Copyright © 2006 European Society of Clinical Infectious Diseases Terms and Conditions