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
{"title":"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。","authors":"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","doi":"10.1128/jcm.00809-24","DOIUrl":null,"url":null,"abstract":"<p><p>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. <i>Treponema pallidum</i> DNA in specimens was detected using quantitative PCR (qPCR), nested PCR, and novel LAMP assays. <i>T. pallidum</i> genotyping was conducted by multi-locus sequence typing (MLST). Of the 163 patients recruited, 98 were diagnosed with syphilis (primary: <i>n</i> = 35; secondary: <i>n</i> = 40; latent: <i>n</i> = 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% (<i>κ</i> coefficient 0.90). The <i>T. pallidum</i> 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 <i>T. pallidum</i> genotype. Dry-LAMP is expected to be helpful in the clinical diagnosis of syphilis.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0080924"},"PeriodicalIF":6.1000,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Prospective evaluation of non-invasive saliva specimens for the diagnosis of syphilis and molecular surveillance of <i>Treponema pallidum</i>.\",\"authors\":\"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\",\"doi\":\"10.1128/jcm.00809-24\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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. <i>Treponema pallidum</i> DNA in specimens was detected using quantitative PCR (qPCR), nested PCR, and novel LAMP assays. <i>T. pallidum</i> genotyping was conducted by multi-locus sequence typing (MLST). Of the 163 patients recruited, 98 were diagnosed with syphilis (primary: <i>n</i> = 35; secondary: <i>n</i> = 40; latent: <i>n</i> = 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% (<i>κ</i> coefficient 0.90). The <i>T. pallidum</i> 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 <i>T. pallidum</i> genotype. Dry-LAMP is expected to be helpful in the clinical diagnosis of syphilis.</p>\",\"PeriodicalId\":15511,\"journal\":{\"name\":\"Journal of Clinical Microbiology\",\"volume\":\" \",\"pages\":\"e0080924\"},\"PeriodicalIF\":6.1000,\"publicationDate\":\"2024-11-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Clinical Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1128/jcm.00809-24\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Clinical Microbiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1128/jcm.00809-24","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
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
Erratum for Ganaie et al., "Discovery and Characterization of Pneumococcal Serogroup 36 Capsule Subtypes, Serotypes 36A and 36B". Multicenter evaluation of Fourier transform infrared (FTIR) spectroscopy as a first-line typing tool for carbapenemase-producing Klebsiella pneumoniae in clinical settings. Yield of solid AFB culture in addition to automated liquid culture for diagnosis of mycobacterial infections. Emergence of terbinafine-resistant Trichophyton indotineae in Ontario, Canada, 2014-2023. Pleural space infection microbiology as assessed using a clinical sequencing-based assay: Fusobacterium nucleatum group, Streptococcus intermedius, and other oral normal microbiota are the most common bacteria identified in community-acquired pleural space infections.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1