Development and evaluation of a test for tuberculosis in live European badgers (Meles meles) based on measurement of gamma interferon mRNA by real-time PCR.

A real-time PCR assay for the measurement of gamma interferon (IFN-gamma) mRNA in European badger (Meles meles) blood cultures was developed. The levels of IFN-gamma mRNA in blood cultures stimulated with either bovine or avian tuberculin or specific mycobacterial antigens were compared with those in a nonstimulated control blood culture as the basis for determining the tuberculosis (TB) status of live badgers. The assay was validated by testing 247 animals for which there were matching data from postmortem examination and culture of tissues. Relative changes in the levels of IFN-gamma mRNA in response to bovine tuberculin and specific antigens were found to be greater among badgers with tissues positive for TB on culture. The test was at its most accurate (87% of test results were correct) by using blood cultures containing bovine tuberculin as the antigen and when the response to avian tuberculin was taken into account by subtracting the avian tuberculin response from the bovine tuberculin response. At a specificity of 90.7%, the test was 70.6% sensitive. At the same specificity, the current serological enzyme-linked immunosorbent assay for TB in badgers was only 53% sensitive. This work demonstrates that measurement of IFN-gamma mRNA by real-time PCR is a valid method for the detection of TB in live badgers and may provide an alternative to the current serological methods of diagnosis, the Brock test. The testing procedure can be completed within 5 h of receipt of the blood culture samples. In addition, the use of a molecular biology-based test offers the potential to fully automate the testing procedure through the use of robotics.

Vibromyographic recording from human muscles with known fibre composition differences.

OBJECTIVE: To determine the relation between the vibromyographic (VMG) frequency characteristics and fibre composition in postural and non-postural human muscle undergoing a standardised voluntary contraction. METHODS: Two human muscles with different fibre compositions [soleus: postural, mainly type I (slow) fibres; biceps brachii: non-postural, mixed type I and II (fast) fibres] were recorded from 18 healthy males isometrically contracting at 50% of their maximum voluntary contraction (MVC). Muscle vibrations were recorded using a contact microphone and the frequency content of the signals calculated using fast fourier transform algorithms. RESULTS: The non-postural biceps brachii showed predominantly bimodal power spectra with significantly increased power in the 10-30 Hz bands (P < 0.01), as compared with soleus recordings which tended to be unimodal, with the majority of power below 10 Hz. CONCLUSIONS: Muscles with a large proportion of type I fibres generate VMG signals which contain an increased percentage of low frequencies as compared to muscles with a mixed population of type I and type II fibres. The VMG appears to be generated, in part at least, by the mechanical twitching of motor units within the muscle; frequency domain analysis of this signal may provide a non-invasive measure of muscle fibre composition.