Gassiep Ian, Glover Matthew, Beecham Mark, Gorman Brian, Page Melissa, Stewart James, Harris Patrick N A
Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Royal Brisbane & Women's Hospital, Queensland, Brisbane, Australia.
Department of Infectious Diseases, Mater Hospital Brisbane, Brisbane, Queensland, Australia.
J Med Microbiol. 2025 Sep;74(9). doi: 10.1099/jmm.0.002074.
Melioidosis is a potentially life-threatening infectious disease. The diagnosis of melioidosis is time-critical due to the organism's intrinsic antimicrobial resistance and requirement for directed therapy. To assess the ability of an automated molecular diagnostic instrument to detect directly from clinical samples. Urine, sputum, swabs and Ashdown's (ASH) broth were spiked with known concentrations of and analysed using an automated PCR platform (Panther Fusion; Hologic) targeting the type III Secretion System (TTS-1) gene. In addition, clinical specimens from patients with confirmed melioidosis were also evaluated. Urine was the clinical sample that demonstrated the lowest limit of detection (LOD), 1.8×10 c.f.u. ml. Compared with dry swabs (LOD: 1.0×10 c.f.u. ml), Amies agar swabs were inferior (LOD: >3.3×10 c.f.u. ml). Inoculation of dry swabs into ASH, with an abbreviated incubation period, did not improve detection. All culture-positive sputum and urine samples from patients with confirmed melioidosis were detected by the PCR method. This study demonstrates the ability of the Panther to directly detect across a range of clinical sample types and estimates the minimum bacterial concentration required for diagnostic detection. The described methodology holds promise for expediting diagnosis and, in turn, enhancing patient outcomes.
类鼻疽是一种潜在的危及生命的传染病。由于该病原体具有内在的抗菌耐药性且需要针对性治疗，类鼻疽的诊断具有时间紧迫性。为评估一种自动化分子诊断仪器直接从临床样本中检测（某病原体）的能力。将已知浓度的（某病原体）添加到尿液、痰液、拭子和阿什当肉汤（ASH）中，并使用靶向III型分泌系统（TTS-1）基因的自动化PCR平台（Panther Fusion；Hologic公司）进行分析。此外，还对确诊类鼻疽患者的临床标本进行了评估。尿液是检测限最低的临床样本，为1.8×10 c.f.u./ml。与干拭子（检测限：1.0×10 c.f.u./ml）相比，阿米斯琼脂拭子的检测效果较差（检测限：>3.3×10 c.f.u./ml）。将干拭子接种到ASH中并缩短培养期，并未提高检测效果。PCR方法检测出了所有确诊类鼻疽患者培养呈阳性的痰液和尿液样本。本研究证明了Panther能够直接检测多种临床样本类型中的（某病原体），并估计了诊断检测所需的最低细菌浓度。所描述的方法有望加快诊断速度，进而改善患者预后。
