Transrenal DNA detection for TB Diagnosis

Coordinator: K. Reither MD, Msc

Background / Purpose:
Tuberculosis (TB) is a global threat to public health. The lack of accurate, rapid, inexpensive and convenient tests for tuberculosis still hinders patient management and disease control. The potential diagnostic capability of cell-free transrenal DNA (Tr-DNA), short DNA fragments which pass the renal barrier, has been demonstrated in infectious diseases, including TB, HIV, and malaria.
The main objectives of the project are to evaluate the potential of transrenal DNA detection in urine specimen for diagnosis of tuberculosis, to optimize and simplify the assay and the specimen preparation, and to explore the suitability of the new technology in a developing world setting. The overall goal will be to obtain the knowledge to develop a robust, accurate and rapid procedure to diagnose TB on the basis of Tr-DNA.

The research project on Tr-DNA is a co-operative venture of 7 participating institutions providing technical and scientific expertise. The project comprises 7 work packages. The project time line is 3 years.
Work package 1 and 2 will take place at the Mbeya Medical Research Center (MMRC / Mbeya, Tanzania) and the University of Zambia (UNZA / Lusaka, Zambia). In this part of the project 900 TB suspected patients and 250 healthy controls will be recruited and followed-up. An established highly sensitive detection assay, using hemi-nested PCR, will be evaluated performing sensitivity and specificity assessment. Furthermore, a bio bank of urine samples from a cohort of patients with symptoms of tuberculosis will be build up.
In the following work packages a small European cohort of TB confirmed patients will be recruited,and a urine bio bank of these clearly defined TB positive patients will be established. The samples will be utilized for optimisation by comparing different extraction procedures for the purification of DNA from urine assessing both the yield of DNA and the removal of nucleic acid amplification inhibitors. In addition, an assessment of potential storage conditions to prevent the degradation of target DNA will be performed. A comprehensive assessment of the biological characteristics of Tr-DNA will be carried out. Moreover, a project database facilitating the interaction and scientific discussion between partners to maximise the final outputs will be established.


European Commission, DG XII

(1) Spaxen Italia Ssrl, Rome, Italy (coordinating partner).
(2) University College London (UCL), London, UK.
(3) University of Zambia (UNZA), Lusaka, Zambia.
(4) Department of Infectious Diseases & Tropical Medicine, Ludwig-Maximilians-University (LMU), Munich, Germany.
(5) The Mbeya Medical Research Center (MMRC), Mbeya, Tanzania.
(6) Istituto Nazionale per le Malattie Infettive L. Spallanzani (INMI), Rome, Italy.
(7) Foundation for Innovative New Diagnostics (FIND), Geneva, Switzerland.

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