Burkholderia pseudomallei is a Gram-negative saprophytic bacillus that causes melioidosis. The infection is endemic in South-East of Asia and Northern Australia. B. pseudomallei has been designated as bioterrorism agent and its manipulation should be done in a biological safety level 3 capability. Workers in laboratories may be accidentally exposed to B. pseudomallei before its identification, with a risk of laboratory-acquired melioidosis. We want to describe a case of melioidosis occurred in our hospital and its management at laboratory. The objective of this article is to provide guidance to microbiologists confronted with a suspicious case of B. pseudomallei on the management of the exposition. We report here a couple of microbiological arguments that can usually guide microbiologists towards presumptive identification of B. pseudomallei. This case report shows the importance of MALDI-TOF MS accurate databases to ensure accurate microbial identification and antibiotic prophylaxis adapted to individuals who were exposed. We also want to underline the importance of developing an effective strategy of prevention against any accidental exposure that can occur in a microbiological laboratory.