对用于诊断梅毒和苍白螺旋体分子监测的非侵入性唾液样本进行前瞻性评估。

IF 6.1 2区 医学 Q1 MICROBIOLOGY Journal of Clinical Microbiology Pub Date : 2024-11-06 DOI:10.1128/jcm.00809-24
Kazuo Imai, Akihiro Sato, Masashi Tanaka, Yuki Ohama, Shu-Ichi Nakayama, Ryuha Omachi, Keita Takeuchi, Norihito Tarumoto, Mieko Tokano, Shigefumi Mesaki, Takuya Maeda, Yukihiro Akeda
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引用次数: 0

摘要

据报道,利用唾液和尿液样本进行梅毒分子检测具有良好的诊断效果;然而,还需要进一步评估其在诊断和分子监测方面的可能应用。此外,开发一种快速、易于操作的梅毒分子检测方法对其在临床环境中的应用也非常重要。我们利用唾液和尿液样本全面评估了两种新型环介导等温扩增(LAMP)检测方法的诊断和监测性能。我们从门诊接受梅毒血清学检测的患者身上采集了唾液、尿液和全血。使用定量 PCR(qPCR)、巢式 PCR 和新型 LAMP 检测法检测标本中的苍白螺旋体 DNA。通过多焦点序列分型(MLST)对苍白螺旋体进行基因分型。在招募的 163 名患者中,98 人被确诊为梅毒患者(原发性:35 人;继发性:40 人;潜伏期:23 人)。在使用唾液和尿液样本进行的所有梅毒患者分子检测中,qPCR 的灵敏度最高,分别为 54.1%和 30.3%。LAMP结合干试剂和DNA粗提取的新方法(Dry-LAMP)在45分钟内的probit检测限为37.4拷贝/反应。唾液中 Dry-LAMP 与 qPCR 的一致率为 95.7%(κ系数 0.90)。通过使用唾液样本进行 MLST,确定了 48 名患者的苍白螺旋体基因型。唾液的分子分析可用作梅毒的辅助诊断检测和苍白螺旋体基因型的分子监测。预计Dry-LAMP有助于梅毒的临床诊断。
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Prospective evaluation of non-invasive saliva specimens for the diagnosis of syphilis and molecular surveillance of Treponema pallidum.

The promising diagnostic performance of molecular testing for syphilis using saliva and urine samples has been reported; however, further evaluation of its possible application for diagnosis and molecular surveillance is required. In addition, the development of a rapid and easy-to-perform molecular test for syphilis is important for its use in the clinical setting. We comprehensively evaluated the diagnostic and surveillance performance of two novel loop-mediated isothermal amplification (LAMP) assays using saliva and urine samples. Saliva, urine, and whole blood were collected from patients who underwent serological testing for syphilis at outpatient clinics. Treponema pallidum DNA in specimens was detected using quantitative PCR (qPCR), nested PCR, and novel LAMP assays. T. pallidum genotyping was conducted by multi-locus sequence typing (MLST). Of the 163 patients recruited, 98 were diagnosed with syphilis (primary: n = 35; secondary: n = 40; latent: n = 23). qPCR showed the highest sensitivity among the molecular tests performed with a sensitivity of 54.1% and 30.3% for all syphilis patients using saliva and urine samples, respectively. A novel method of LAMP combined with dry reagents and crude DNA extraction (Dry-LAMP) showed a probit detection limit of 37.4 copies/reaction within 45 min. The agreement rate between Dry-LAMP and qPCR for saliva was 95.7% (κ coefficient 0.90). The T. pallidum genotype was identified in 48 patients by MLST using saliva samples. Molecular analysis of saliva could be used as a supplementary diagnostic test for syphilis and molecular surveillance of the T. pallidum genotype. Dry-LAMP is expected to be helpful in the clinical diagnosis of syphilis.

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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
期刊最新文献
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