IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v13y2022i1d10.1038_s41467-022-32250-y.html
   My bibliography  Save this article

A suite of PCR-LwCas13a assays for detection and genotyping of Treponema pallidum in clinical samples

Author

Listed:
  • Wentao Chen

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Hao Luo

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Lihong Zeng

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Yuying Pan

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Jonathan B. Parr

    (University of North Carolina)

  • Yinbo Jiang

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Clark H. Cunningham

    (University of North Carolina)

  • Kelly L. Hawley

    (Connecticut Children’s
    UConn Health
    UConn Health)

  • Justin D. Radolf

    (UConn Health
    UConn Health
    UConn Health
    UConn Health)

  • Wujian Ke

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Jiangli Ou

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Jianjiang Yang

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Bin Yang

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

  • Heping Zheng

    (Southern Medical University
    Guangzhou Key Laboratory for Sexually Transmitted Diseases Control)

Abstract

The performance of commonly used assays for diagnosis of syphilis varies considerably depending on stage of infection and sample type. In response to the need for improved syphilis diagnostics, we develop assays that pair PCR pre-amplification of the tpp47 gene of Treponema pallidum subsp. pallidum with CRISPR-LwCas13a. The PCR-LwCas13a assay achieves an order of magnitude better analytical sensitivity than real-time PCR with equivalent specificity. When applied to a panel of 216 biological specimens, including 135 clinically confirmed primary and secondary syphilis samples, the PCR-LwCas13a assay demonstrates 93.3% clinical sensitivity and 100% specificity, outperforming tpp47 real-time PCR and rabbit-infectivity testing. We further adapt this approach to distinguish Treponema pallidum subsp. pallidum lineages and identify genetic markers of macrolide resistance. Our study demonstrates the potential of CRISPR-based approaches to improve diagnosis and epidemiological surveillance of syphilis.

Suggested Citation

  • Wentao Chen & Hao Luo & Lihong Zeng & Yuying Pan & Jonathan B. Parr & Yinbo Jiang & Clark H. Cunningham & Kelly L. Hawley & Justin D. Radolf & Wujian Ke & Jiangli Ou & Jianjiang Yang & Bin Yang & Hepi, 2022. "A suite of PCR-LwCas13a assays for detection and genotyping of Treponema pallidum in clinical samples," Nature Communications, Nature, vol. 13(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-32250-y
    DOI: 10.1038/s41467-022-32250-y
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-022-32250-y
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-022-32250-y?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. Simant Dube & Jian Qin & Ramesh Ramakrishnan, 2008. "Mathematical Analysis of Copy Number Variation in a DNA Sample Using Digital PCR on a Nanofluidic Device," PLOS ONE, Public Library of Science, vol. 3(8), pages 1-9, August.
    2. Mathew A. Beale & Michael Marks & Sharon K. Sahi & Lauren C. Tantalo & Achyuta V. Nori & Patrick French & Sheila A. Lukehart & Christina M. Marra & Nicholas R. Thomson, 2019. "Genomic epidemiology of syphilis reveals independent emergence of macrolide resistance across multiple circulating lineages," Nature Communications, Nature, vol. 10(1), pages 1-9, December.
    Full references (including those not matched with items on IDEAS)

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Davide Cardinale & Maria Tafuro & Andrea Mancusi & Santa Girardi & Federico Capuano & Yolande Thérèse Rose Proroga & Federica Corrado & Jacopo Luigi D’Auria & Annachiara Coppola & Giuseppe Rofrano & P, 2022. "Sponge Whirl-Pak Sampling Method and Droplet Digital RT-PCR Assay for Monitoring of SARS-CoV-2 on Surfaces in Public and Working Environments," IJERPH, MDPI, vol. 19(10), pages 1-14, May.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-32250-y. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.