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There is an urgent need for simple, accurate, inexpensive and rapid diagnostic tests for TB. Thin-Layer Agar (TLA) has been proposed as an easily implemented technique of culturing and identifying Mycobacteria and has been shown to be rapid and at least equally sensitive as conventional methods [5]. Most of the studies on TLA come from developed countries [6-8] or from Latin America [5, 9-12]. Many included mixed samples [6-8, 11-13], some come from high HIV prevalence settings [14]. We aimed to compare the performance of culture of sputum on TLA with culture on L?wenstein�CJensen (LJ) medium in terms of accuracy, contamination rate and turnaround time, for diagnosing pulmonary TB, in a setting with high TB incidence, low HIV prevalence, and limited resources. The study took place in Jogjakarta municipality, Indonesia, which has a population of around 500?000, an estimated BGJ398 clinical trial TB incidence of 63/100?000 [15] and 1.9% HIV prevalence among TB patients [16]. The National TB Programme relies on nine hospitals, 18 health centres and two chest clinics. Four health centres perform smear microscopy. The chest clinics perform smear microscopy, chest X-ray and offer voluntary HIV counselling and testing; they are run by general practitioners and they are regularly visited by chest specialists. The clinics accept patients referred from health centres or self-referred. The Microbiology Laboratory, Faculty of Medicine, Gadjah Mada University is a biosafety level II plus TB Fluconazole diagnostic Paclitaxel referral laboratory performing microscopy, Mycobacterium culture using LJ and drug susceptibility testing using proportional methods. It has a biosafety cabinet class two type B2. The study was carried out in the two chest clinics and in the Microbiology Laboratory of Gadjah Mada University and it was endorsed by the Indonesian National TB Programme. Participants were included prospectively. All new patients with suspected TB presenting from July 2010 to July 2011 to the chest clinics were eligible. Following national guidelines [17], they were individuals with cough lasting more than 2?weeks, never treated for TB. Exclusion criteria were age