PCR Techniques of Dried Sputum Using a Spotcard
PCR Techniques of Dried Sputum Using a Spotcard
Molecular testing for mutations in M. tuberculosis genes associated to resistance of anti
 tuberculosis (TB) drugs is already part of standard laboratory TB diagnostic.

 This implicates earlier knowledge of possible resistance and thus prevents unnecessary
 treatment and the chance of treatment failure or treatment related toxicity.

 The molecular laboratory diagnostics is widely spread in high income, low TB endemic
 countries. However, the low income countries lack widespread facilities to test for
 susceptibility, either genotypic or phenotypic. Performing molecular diagnostics on sputum
 collected with a spot card could improve accessibility to molecular testing.

 This study examines if sputum collected and put on spot cards could be used for multiple
 molecular tests for the detection, identification and susceptibility prediction of TB. This
 means that DNA extraction of the sputum from the spot card should be feasible.

 The study is a pilot study with adult patients of the tuberculosis department of University
 Medical Centre Groningen (UMCG) Beatrixoord, Haren as subjects. The sputum produced will be
 collected, dried on spot cards, and DNA extraction from the card will be tested. If molecular
 detection is positive for the tuberculosis bacteria additional tests will be performed. Based
 on the present/absent of mutations in the genes associated to resistance susceptibility can
 be preditec, different molecular techniques will be performed to identify possible mutations.
 Furthermore, sputum will be collected as patients produce so. Sputum samples with low
 bacterial load can be tested as well and can test the sensitivity of the procedure.

 Lastly, techniques like RNA detection will be tested to identify the bacterial load. This can
 be done if more than one sample of patients were collected.

 Subjects will be selected on age, participation in standard TDM and drug use. Demographic
 parameters will be analysed. Sputum samples will be taken twice a week (on Tuesday and
 Friday).
Inclusion Criteria:

 -  Patients is 18 years or older

 -  Pulmonary TB confirmed by culture or molecular test

 -  Producing sputum

 -  Signed informed consent

 Exclusion Criteria:

 -  Extrapulmonary TB

 -  Pulmonary TB without production of sputum