Performance of the 47-Kilodalton Membrane Protein versus DNA Polymerase I Genes for Detection of Treponema pallidum by PCR in Ulcers

Treponema pallidumPCR (Tp-PCR) is a direct diagnostic method for primary and secondary syphilis, but there is no recommendation regarding the best choice of target gene. In this study, we sequentially tested 272 specimens from patients with sexually transmitted ulcers usingTp-PCR targeting thetpp47and thenpolAgenes. The two methods showed similar accuracies and an almost-perfect agreement.

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Treponema pallidumNucleic Acid Amplification Testing To Augment Syphilis Screening among Men Who Have Sex with Men

Journal of Clinical Microbiology ◽

10.1128/jcm.00572-19 ◽

2019 ◽

Vol 57 (8) ◽

Cited By ~ 1

Author(s):

Matthew Golden ◽

Meghan O’Donnell ◽

Sheila Lukehart ◽

Paul Swenson ◽

Paul Hovey ◽

...

Keyword(s):

Transmitted Disease ◽

Treponema Pallidum ◽

Secondary Syphilis ◽

23S Rrna ◽

Serological Testing ◽

Content Type ◽

Sexually Transmitted ◽

Syphilis Screening ◽

Sex With Men ◽

Latent Syphilis

ABSTRACTSyphilis rates in much of the world are now at their highest levels in almost three decades, and new approaches to controlling syphilis, including diagnostic tests with shorter window periods, are urgently needed. We compared the sensitivity of syphilis serological testing using the rapid plasma reagin (RPR) test with that of the combination of serological testing and an experimental 23S rRNATreponema pallidumreal-time transcription-mediated amplification (TMA) assay performed on rectal and pharyngeal mucosal swabs.T. pallidumPCR assays for thetpp47gene were performed on all TMA-positive specimens, as well as specimens from 20 randomly selected TMA-negative men. A total of 545 men who have sex with men (MSM) who were seen in a sexually transmitted disease clinic provided 506 pharyngeal specimens and 410 rectal specimens with valid TMA results. Twenty-two men (4%) were diagnosed with syphilis on the basis of positive RPR test results and clinical diagnoses, including 3 men with primary infections, 8 with secondary syphilis, 9 with early latent syphilis, 1 with late latent syphilis, and 1 with an unstaged infection. Two additional men were diagnosed based on positive rectal mucosal TMA assay results alone, and both also tested positive by PCR assay. At least 1 specimen was TMA positive for 12 of 24 men with syphilis (sensitivity, 50% [95% confidence interval [CI], 29 to 71%]). RPR testing and clinical diagnosis were 92% sensitive (95% CI, 73 to 99%) in identifying infected men. Combining mucosal TMA testing and serological testing may increase the sensitivity of syphilis screening in high-risk populations.

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Comparison of Diagnostic Accuracy of PCR Targeting the 47-Kilodalton Protein Membrane Gene of Treponema pallidum and PCR Targeting the DNA Polymerase I Gene: Systematic Review and Meta-analysis

Journal of Clinical Microbiology ◽

10.1128/jcm.01619-15 ◽

2015 ◽

Vol 53 (11) ◽

pp. 3522-3529 ◽

Cited By ~ 14

Author(s):

Angèle Gayet-Ageron ◽

Christophe Combescure ◽

Stephan Lautenschlager ◽

Béatrice Ninet ◽

Thomas V. Perneger

Keyword(s):

Meta Analysis ◽

Treponema Pallidum ◽

Quality Criteria ◽

Secondary Syphilis ◽

Bibliographic Databases ◽

Case Definitions ◽

Content Type ◽

Early Syphilis ◽

Similar Accuracy ◽

Pcr Targeting

Treponema pallidumPCR (Tp-PCR) testing now is recommended as a valid tool for the diagnosis of primary or secondary syphilis. The objectives were to systematically review and determine the optimal specific target gene to be used forTp-PCR. Comparisons of the performance of the two main targets aretpp47andpolAgenes were done using meta-analysis. Three electronic bibliographic databases, representing abstract books from five conferences specialized in infectious diseases from January 1990 to March 2015, were searched. Search keywords included (“syphilis” OR “Treponema pallidum” OR “neurosyphilis”) AND (“PCR” OR “PCR” OR “molecular amplification”). We included diagnostic studies assessing the performance ofTp-PCR targetingtpp47(tpp47-Tp-PCR) or thepolAgene (polA-Tp-PCR) in ulcers from early syphilis. All studies were assessed against quality criteria using the QUADAS-2 tool. Of 37 studies identified, 62.2% were judged at low risk of bias or applicability. Most used the U.S. Centers for Disease Control and Prevention (CDC) case definitions for primary or secondary (early) syphilis (89.2%;n= 33); 15 (40.5%) used darkfield microscopy (DFM). We did not find differences in sensitivity and specificity between the twoTp-PCR methods in the subgroup of studies using adequate reference tests. Among studies using DFM as the reference test, sensitivities were 79.8% (95% confidence intervals [CI], 72.7 to 85.4%) and 71.4% (46.0 to 88.0%) fortpp47-Tp-PCR andpolA-Tp-PCR (P= 0.217), respectively; respective specificities were 95.3% (93.5 to 96.6%) and 93.7% (91.8 to 95.2%) (P= 0.304). Our findings suggest that the twoTp-PCR methods have similar accuracy and could be used interchangeably.

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Molecular cloning of a gene (polA) coding for an unusual DNA polymerase I from Treponema pallidum

Journal of Medical Microbiology ◽

10.1099/0022-1317-49-7-657 ◽

2000 ◽

Vol 49 (7) ◽

pp. 657-667 ◽

Cited By ~ 7

Author(s):

BERTA RODES ◽

HSI LIU ◽

ROBERT GEORGE ◽

STEVEN JOHNSON ◽

BRET STEINER

Keyword(s):

Molecular Cloning ◽

Dna Polymerase ◽

Treponema Pallidum ◽

Dna Polymerase I ◽

Polymerase I

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A New Specimen for Syphilis Diagnosis: Evidence by High Loads of Treponema pallidum DNA in Saliva

Clinical Infectious Diseases ◽

10.1093/cid/ciaa1613 ◽

2020 ◽

Author(s):

Cuini Wang ◽

Zhixiang Hu ◽

Xin Zheng ◽

Meiping Ye ◽

Chunjie Liao ◽

...

Keyword(s):

Treponema Pallidum ◽

Digital Pcr ◽

Secondary Syphilis ◽

Clearance Time ◽

Chain Reaction ◽

Nested Polymerase Chain Reaction ◽

Detection Rates ◽

Polymerase Chain ◽

Pcr Targeting ◽

Latent Syphilis

Abstract Background DNA from many pathogens can be detected in saliva. However, the presence and quantity of Treponema pallidum DNA in patients with syphilis in saliva is unknown. Methods 234 patients with syphilis with different stages and 30 volunteers were enrolled. Paired saliva and plasma samples were collected from all participants. Consecutive saliva samples from 9 patients were collected every 4 hours following treatment. Treponema pallidum DNA in samples was determined by nested polymerase chain reaction (PCR) and droplet digital PCR targeting polA and Tpp47. Results Treponema pallidum DNA detection rates in saliva and plasma were 31.0% (9/29) and 51.7% (15/29) in primary syphilis (P = .11), 87.5% (63/72) and 61.1% (44/72) in secondary syphilis (P < .001), 25.6% (21/82) and 8.5% (7/82) in latent syphilis (P = .004), and 21.6% (11/51) and 5.9% (3/51) in symptomatic neurosyphilis (P = .021), respectively. Median (range) loads of Tpp47 and polA in saliva were 627 (0–101 200) and 726 (0–117 260) copies/mL, respectively, for patients with syphilis. In plasma, however, loads of Tpp47 and polA were low: medians (range) of 0 (0–149.6) and 0 (0–176) copies/mL, respectively. Loads of T. pallidum DNA in saliva during treatment fluctuated downward; the clearance time was positively correlated with the loads of T. pallidum DNA before treatment. Conclusions Collection of saliva is noninvasive and convenient. The high loads of T. pallidum DNA in saliva and reduction after treatment indicated that saliva can be not only a diagnostic fluid for syphilis but also an indicator of therapeutic effectiveness.

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High-Efficiency Scarless Genetic Modification in Escherichia coli by Using Lambda Red Recombination and I-SceI Cleavage

Applied and Environmental Microbiology ◽

10.1128/aem.00313-14 ◽

2014 ◽

Vol 80 (13) ◽

pp. 3826-3834 ◽

Cited By ~ 44

Author(s):

Junjie Yang ◽

Bingbing Sun ◽

He Huang ◽

Yu Jiang ◽

Liuyang Diao ◽

...

Keyword(s):

Escherichia Coli ◽

Genetic Modification ◽

Target Gene ◽

Donor Plasmid ◽

Content Type ◽

Recognition Sites ◽

Genetic Modifications ◽

Pcr Targeting ◽

Λ Red ◽

Red Recombination

ABSTRACTGenetic modifications of bacterial chromosomes are important for both fundamental and applied research. In this study, we developed an efficient, easy-to-use system for genetic modification of theEscherichia colichromosome, a two-plasmid method involving lambda Red (λ-Red) recombination and I-SceI cleavage. An intermediate strain is generated by integration of a resistance marker gene(s) and I-SceI recognition sites in or near the target gene locus, using λ-Red PCR targeting. The intermediate strain is transformed with a donor plasmid carrying the target gene fragment with the desired modification flanked by I-SceI recognition sites, together with a bifunctional helper plasmid for λ-Red recombination and I-SceI endonuclease. I-SceI cleavage of the chromosome and the donor plasmid allows λ-Red recombination between chromosomal breaks and linear double-stranded DNA from the donor plasmid. Genetic modifications are introduced into the chromosome, and the placement of the I-SceI sites determines the nature of the recombination and the modification. This method was successfully used forcadAknockout,gdhAknock-in, seamless deletion ofpepD, site-directed mutagenesis of the essentialmetKgene, and replacement ofmetKwith theRickettsiaS-adenosylmethionine transporter gene. This effective method can be used with both essential and nonessential gene modifications and will benefit basic and applied genetic research.

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Evaluation of the HISCL Anti-Treponema pallidum Assay as a Screening Test for Syphilis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00116-15 ◽

2015 ◽

Vol 22 (7) ◽

pp. 817-822 ◽

Cited By ~ 6

Author(s):

Jingna An ◽

Qixia Chen ◽

Qianqian Liu ◽

Chenli Rao ◽

Dongdong Li ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Screening Test ◽

Treponema Pallidum ◽

High Volume ◽

Laboratory Diagnosis ◽

Ease Of Use ◽

Perfect Agreement ◽

Content Type ◽

Syphilis Screening ◽

Testing Algorithms

ABSTRACTThe resurgence of syphilis in recent years has become a serious threat to public health worldwide, and the serological detection of specific antibodies againstTreponema pallidumremains the most reliable method for laboratory diagnosis of syphilis. This study examined the performance of the recently launched HISCL anti-Treponema pallidum(anti-TP) assay as a screening test for syphilis in a high-volume laboratory. The HISCL anti-TP assay was tested in 300 preselected syphilis-positive samples, 704 fresh syphilis-negative samples, 48 preselected potentially interfering samples, and 30 “borderline” samples and was compared head to head with the commercially available Lumipulse G TP-N. In this study, the HISCL anti-TP assay was in perfect agreement with the applied testing algorithms with an overall agreement of 100%, comparable to that of Lumipulse G TP-N (99.63%). The sensitivity and specificity of the HISCL anti-TP assay were 100% (95% confidence interval [CI], 98.42% to 100%) and 100% (95% CI, 99.37% to 100%), respectively. Considering the excellent ease of use and automation, high throughput, and its favorable sensitivity and specificity, the HISCL anti-TP assay may represent a new choice for syphilis screening in high-volume laboratories.

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New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene

Journal of Clinical Microbiology ◽

10.1128/jcm.39.5.1941-1946.2001 ◽

2001 ◽

Vol 39 (5) ◽

pp. 1941-1946 ◽

Cited By ~ 129

Author(s):

H. Liu ◽

B. Rodes ◽

C.-Y. Chen ◽

B. Steiner

Keyword(s):

Dna Polymerase ◽

Rational Design ◽

Treponema Pallidum ◽

Dna Polymerase I ◽

Clinical Specimens ◽

Pcr Method ◽

Polymerase I ◽

I Gene

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Imunopatogenesis Treponema pallidum dan Pemeriksaan Serologi

Jurnal Kesehatan Andalas ◽

10.25077/jka.v3i3.203 ◽

2014 ◽

Vol 3 (3) ◽

Author(s):

Efrida Efrida ◽

Elvinawaty Elvinawaty

Keyword(s):

Clinical Symptoms ◽

Early Stage ◽

Transmitted Disease ◽

Treponema Pallidum ◽

Secondary Syphilis ◽

Major Health Problem ◽

Sexually Transmitted ◽

Tertiary Syphilis ◽

History Of ◽

Latent Syphilis

AbstrakSifilis adalah penyakit menular seksual yang sangat infeksius, disebabkan oleh bakteri berbentuk spiral, Treponema pallidum subspesies pallidum. Penyebaran sifilis di dunia telah menjadi masalah kesehatan yang besar dengan jumlah kasus 12 juta pertahun. Infeksi sifilis dibagi menjadi sifilis stadium dini dan lanjut. Sifilis stadium dini terbagi menjadi sifilis primer, sekunder, dan laten dini. Sifilis stadium lanjut termasuk sifilis tersier (gumatous, sifilis kardiovaskular dan neurosifilis) serta sifilis laten lanjut. Sifilis primer didiagnosis berdasarkan gejala klinis ditemukannya satu atau lebih chancre (ulser). Sifilis sekunder ditandai dengan ditemukannya lesi mukokutaneus yang terlokalisir atau difus dengan limfadenopati. Sifilis laten tanpa gejala klinis sifilis dengan pemeriksaan nontreponemal dan treponemal reaktif, riwayat terapi sifilis dengan titer uji nontreponemal yang meningkat dibandingkan dengan hasil titer nontreponemal sebelumnya. Sifilis tersier ditemukan guma dengan pemeriksaan treponemal reaktif, sekitar 30% dengan uji nontreponemal yang tidak reaktifKata kunci: sifilis, Treponema pallidum, serologiAbstractSyphilis is a sexually transmitted disease that is highly infectious, caused by a spiral -shaped bacterium, Treponema pallidum subspecies pallidum. The spread of syphilis in the world has become a major health problem and the common, the number of 12 million cases per year. Infectious syphilis is divided into early and late-stage syphilis. Early-stage syphilis is divided into primary, secondary, and early latent. Advanced stage of syphilis include tertiary syphilis (gumatous, cardiovascular syphilis, and neurosyphilis) and late latent syphilis. Primary syphilis is diagnosed by clinical symptoms of the discovery of one or more chancre (ulcer). Secondary syphilis is characterized by the finding of localized mucocutaneous lesions or with diffuse lymphadenopathy. Latent syphilis without clinical symptoms of syphilis with a nontreponemal and treponemal reactive examination, history of syphilis therapy in nontreponemal test titer increased compared with the results of previous nontreponemal titers. Tertiary syphilis is found guma with reactive treponemal examination, approximately 30% of the non- reactive nontreponemal testKeywords: syphilis, Treponema pallidum, serologi

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Exploring the Ways of “The Great Imitator”: A Case Report of Syphilitic Hepatitis

GE Portuguese Journal of Gastroenterology ◽

10.1159/000516944 ◽

2021 ◽

pp. 1-4

Author(s):

João A. Cunha Neves ◽

Joana Roseira ◽

Helena Tavares de Sousa ◽

Rui Machado

Keyword(s):

Immune Responses ◽

Venereal Disease ◽

Treponema Pallidum ◽

Maculopapular Rash ◽

Liver Function Tests ◽

Secondary Syphilis ◽

Penicillin G ◽

Sexually Transmitted ◽

Patients At Risk ◽

Mixed Pattern

Introduction: Syphilis is a chronic infection caused by Treponema pallidum. Manifestations of this disease are vast, and syphilitic hepatitis is a rarely depicted form of secondary syphilis. Case Presentation: We report the case of a 63-year-old man with worsening jaundice, maculopapular rash and perianal discomfort. Proctological examination with anoscopy revealed a perianal gray/white area with millimetric pale granules along the anal canal. Liver function tests showed a mixed pattern. Venereal Disease Research Laboratory, T. pallidum hemagglutination assay and IgM fluorescent treponemal antibody absorbance were positive. The patient was successfully treated with a single dose of penicillin G. Discussion/Conclusion: Syphilitic hepatitis is scarcely reported in the literature. Secondary syphilis with mild hepatitis rarely leads to hepatic cytolysis and jaundice. Many signs of secondary syphilis including syphilitic hepatitis may be linked to immune responses initiated during early infection. Over the past decades, evidence has emerged on the importance of innate and adaptive cellular immune responses in the immunopathogenesis of syphilis. This report raises awareness to a clinical entity that should be considered in patients at risk for sexually transmitted diseases, who present with intestinal discomfort or liver dysfunction, as it is a treatable and fully reversible condition.

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