Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 21, Number 3—March 2015
Dispatch

Noninvasive Test for Tuberculosis Detection among Primates

Tiffany M. WolfComments to Author , Lawrence Mugisha, Fernanda Miyagaki Shoyama, Melanie J. O’Malley, JoAnne L. Flynn, Benon Asiimwe, Dominic A. Travis, Randall S. Singer, and Srinand Sreevatsan
Author affiliations: Minnesota Zoological Gardens, Apple Valley, Minnesota, USA (T.M. Wolf); University of Minnesota, St. Paul, Minnesota, USA (T.M. Wolf, L. Mugisha, F.M. Shoyama, D.A. Travis, R.S. Singer, S. Sreevatsan); Conservation & Ecosystem Health Alliance, Kampala, Uganda (L. Mugisha); Makerere University, Kampala (L. Mugisha, B. Asiimwe); University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA (M.J. O’Malley, J.L. Flynn)

Main Article

Table 2

Fecal IS6110 conventional and real-time PCR master mixes and reaction conditions for investigation of noninvasive tuberculosis detection in primates

PCR type Primers, 5′ → 3′ Master mix Reaction conditions
Conventional
 Forward: TTCAGGTCGAGTACGCCTTC Reverse: CGAACTCAAGGAGCACATCA
 12.5 μL HotStarTaq Master Mix:* 8 μL RNase-free water,* 0.4 μM of each primer, 1.25 μL DMSO, 0.25 μL 1% BSA, 1 μL DNA template. Total volume 25 μL
 95°C for 15 min/DNA polymerase activation; 40 cycles: 94°C for 30 s/denaturation, 56°C for 30 s/annealing, 72°C for 1 min/extension. Termination at 72°C for 10 min
Real-time Forward: AGAAGGCGTACTCGACCTGA Reverse: CCGGATCGATGTGTACTGAG LightCycler 480 Probes Master:† 0.2 mM of each primer, 0.2 mM of the FAM-labeled IS6110 probe,† 5 μL DNA template. Total volume 25 μL 95°C for 5 min; 45 cycles: 95°C for 10 s/denaturation; 50°C for 30 s/annealing; 72°C for 1 s/extension. Termination at 65°C–95°C at 2.2°C/s/melting curve analysis

*QIAGEN, Inc., Valencia, CA, USA.
†Roche, Indianapolis, IN, USA

Main Article

Page created: February 18, 2015
Page updated: February 18, 2015
Page reviewed: February 18, 2015
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
file_external