Mycoplasma pneumoniae is a strong infectious pathogen that may cause severe respiratory infections. Since this pathogen may possess a latent period after infection, which sometimes leads to misdiagnosis by traditional diagnosis methods, the establishment of a rapid and sensitive diagnostic method is crucial for transmission prevention and timely treatment. Herein, a novel detection method was established for M. pneumoniae detection. The method, which improves upon a denaturation bubble-mediated strand exchange amplification (SEA) that we developed in 2016, is called accelerated SEA (ASEA). The established ASEA achieved detection of 1% M. pneumoniae genomic DNA in a DNA mixture from multiple pathogens, and the limit of detection (LOD) of ASEA was as low as 1.0 × 10⁻¹⁷ M (approximately 6.0 × 10³ copies/mL). Considering that the threshold of an asymptomatic carriage is normally recommended as 1.0 × 10⁴ copies/mL, this method was able to satisfy the requirement for practical diagnosis of M. pneumoniae. Moreover, the detection process was finished within 20.4 min, significantly shorter than real-time PCR and SEA. Furthermore, ASEA exhibited excellent performance in clinical specimen analysis, with sensitivity and specificity of 96.2% and 100%, respectively, compared with the “gold standard” real-time PCR. More importantly, similar to real-time PCR, ASEA requires only one pair of primers and ordinary commercial polymerase, and can be carried out using a conventional fluorescence real-time PCR instrument, which makes this method low-cost and easy to accomplish. Therefore, ASEA has the potential for wide use in the rapid detection of M. pneumoniae or other pathogens in large numbers of specimens.

肺炎支原体是一种强传染性病原体，可引起严重的呼吸道感染。由于该病原体在感染后可能具有潜伏期，有时会导致传统诊断方法误诊，因此建立快速灵敏的诊断方法对于预防传播和及时治疗至关重要。在此，建立了一种用于检测肺炎支原体的新检测方法。该方法改进了我们在 2016 年开发的变性气泡介导的链交换扩增 (SEA)，称为加速 SEA (ASEA)。建立的ASEA实现了对多种病原体DNA混合物中1%肺炎支原体基因组DNA的检测，ASEA的检测限（LOD）低至1.0×10^-17  M（约 6.0 × 10 ^{3 个}拷贝/mL）。考虑到无症状携带者的阈值通常推荐为1.0×10 ⁴拷贝/mL，该方法能够满足肺炎支原体的实际诊断要求。. 此外，检测过程在 20.4 分钟内完成，明显短于实时 PCR 和 SEA。此外，ASEA 在临床标本分析方面表现出优异的性能，与“金标准”实时 PCR 相比，灵敏度和特异性分别为 96.2% 和 100%。更重要的是，与实时荧光定量PCR类似，ASEA只需要一对引物和普通的商业聚合酶，并且可以使用常规的荧光实时荧光定量PCR仪器进行，这使得该方法成本低且易于完成。因此，ASEA 具有广泛用于快速检测大量标本中的肺炎支原体或其他病原体的潜力。