The present invention relates to an isolated polynucleotide for detecting and identifying Burkholderia pseudomallei in a biological sample, comprising a nucleotide sequence set forth in SEQ ID NO: 1 or the complementary sequence thereof. It also relates to a method for detecting and identifying Burkholderia pseudomallei in a biological sample, comprising the steps of constructing a labelledprobe having a nucleotide sequence set forth in SEQ ID NO: 1 or the complementary sequence thereof; pretreating a biological sample; hybridizing the probe onto the pretreated biological sample in situ; and visualizing the hybridized probe via a chromogenic imaging system.