Krueger Trinity, Kadonsky Grace, Thelen Elizabeth, Zapp Amanda, Moore Josephine, Muslu Ahmet, Pillay Allan, Stafford Irene A, Munson Erik
1Department of Medical Laboratory Science, Marquette University, Milwaukee, Wisconsin, USA.
Division of STD Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
J Clin Microbiol. 2025 Aug 13;63(8):e0038825. doi: 10.1128/jcm.00388-25. Epub 2025 Jun 20.
Preliminary experimentation has suggested that a research-use-only real-time transcription-mediated amplification assay for (RUO TMA) yields instances of nucleic acid detection that coincide with non-treponemal serology in men who have sex with men (MSM) at increased risk for sexually transmitted infection. To further characterize the specificity of RUO TMA testing, 3,586 rectal swab specimens reported as "not detected" by the assay generated a mean endpoint FAM fluorescence of 637.5 units. Introduction of , , and nucleic acid into matrices generated mean endpoint FAM fluorescence ranging from 620.7 to 633.5 units ( ≥ 0.15 versus control). Introduction of lubricant to a pooled rectal swab matrix did not result in elevated FAM fluorescence (mean endpoint value 628.3 units [95% CI 612.0, 644.6]; = 0.67). Introduction of talcum powder, urine, seminal fluid, and blood also failed to generate increased FAM fluorescence ( ≥ 0.20). Analytic sensitivity assessment was measured by serial 10-fold dilution of whole organism or 23S rRNA transcript in specimen transport medium or pooled rectal swab matrix and interrogation by the assay. Probit analysis estimated sensitivity (95% detection) of RUO TMA at 421-5,707 transcript copies/mL and 9-48 cells/mL, depending on dilution matrix. These results support RUO TMA as a highly sensitive method for detection that is not impacted by potentially cross-reactive organisms or interfering substances and may have adjunctive diagnosis capability for syphilis in MSM.
Research-use-only transcription-mediated amplification (RUO TMA) has the potential to improve laboratory diagnosis of syphilis, particularly in patients at increased risk for sexually transmitted infection. The high analytic sensitivity and lack of cross-reactivity of the assay can facilitate other laboratories exploring the use of the test in a research setting.
初步实验表明，一种仅供研究使用的实时转录介导扩增检测方法（RUO TMA）在性传播感染风险增加的男男性行为者（MSM）中，其核酸检测结果与非梅毒螺旋体血清学结果相符。为了进一步表征RUO TMA检测的特异性，该检测报告为“未检测到”的3586份直肠拭子标本产生的平均终点FAM荧光为637.5单位。将 、 和 核酸引入基质后，产生的平均终点FAM荧光范围为620.7至633.5单位（与对照相比，P≥0.15）。向混合直肠拭子基质中添加润滑剂不会导致FAM荧光升高（平均终点值628.3单位[95%CI 612.0, 644.6]；P = 0.67）。添加滑石粉、尿液、精液和血液也未能产生更高的FAM荧光（P≥0.20）。通过在标本运输培养基或混合直肠拭子基质中对 全菌或 23S rRNA转录本进行连续10倍稀释并进行该检测来评估分析灵敏度。概率分析估计，根据稀释基质的不同，RUO TMA的灵敏度（95%检测率）在421 - 5707个转录本拷贝/mL和9 - 48个细胞/mL之间。这些结果支持RUO TMA作为一种高度灵敏的 检测方法，不受潜在交叉反应性生物体或干扰物质的影响，可能对MSM梅毒具有辅助诊断能力。
仅供研究使用的 转录介导扩增（RUO TMA）有潜力改善梅毒的实验室诊断，特别是在性传播感染风险增加的患者中。该检测方法的高分析灵敏度和缺乏交叉反应性有助于其他实验室在研究环境中探索使用该检测方法。
