Analytical performance characteristics of a new transcription-mediated amplification assay for Treponema pallidum

This study evaluated the performance characteristics of a new research use transcription-mediated amplification (TMA) assay for detection of ribosomal RNA from Treponema pallidum (Tp). Analytical sensitivity determined using dark field quantitated Tp was 1.4 organisms/ml (95% CI: 0.7-6.33). Dilution of Tp IVT RNA in STM yielded 100% positivity (n=3) at 10 copies/ml (4 copies/reaction). Analytical specificity testing of non-target microorganisms (N=59), including the closely related non-syphilis treponemes T. denticola and T. phagedenis, yielded 0% positivity. TMA testing of mucosal swab specimens collected from men who have sex with men (MSM) attending a sexually transmitted disease clinic yielded 1.8% (17/944) TMA positive results. A collection of 56 serum specimens obtained from a separate cohort of patients with known rapid plasma reagin (RPR) status and syphilis clinical diagnosis were 19.6% (11/56) TMA positive overall, 29.7% (11/37) positive among subjects with syphilis diagnosis, including 8 (36.3%) of 22 persons with primary or secondary syphilis, 2 (20%) of 10 persons with early latent syphilis and 1 (20%) of 5 persons with late latent or unstaged syphilis. None (0%) of 18 RPR positive sera from patients with a history of treated syphilis was TMA positive. These results show TMA is an analytically sensitive and specific method for detection of Tp rRNA and is compatible with serum specimens in addition to pharyngeal and rectal mucocutaneous swab specimens. Automated real time TMA testing for T. pallidum may be useful as an adjunctive method with serology for screening and diagnostic testing of selected patient populations for syphilis.

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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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Laboratory Examination of Syphilis

BioScientia Medicina ◽

10.32539/bsm.v5i3.339 ◽

2021 ◽

Vol 5 (3) ◽

pp. 722-741

Author(s):

M. Izazi Hari Purwoko ◽

Mutia Devi ◽

Suroso Adi Nugroho ◽

Fitriani Fitriani ◽

Raden Pamudji ◽

...

Keyword(s):

Venereal Disease ◽

Transmitted Disease ◽

Treponema Pallidum ◽

Clinical Manifestations ◽

Rapid Test ◽

Dark Field ◽

Morphological Observation ◽

Serologic Test ◽

Sexually Transmitted ◽

Antibody Absorption

Syphilis, is sexually transmitted disease caused by spirochete Treponema pallidum subsp.pallidum. It have many diverse clinical manifestations that occur in distinct stages. Early diagnosis and management are the main things to prevent transmission and complication. Direct test or morphological observation is the definitive diagnosis of syphilis. This can be done through animal inoculation test, dark field microscopy, direct fluorescence antibody (DFA), and nucleid acid amplification test (NAAT). While the indirect test is a nontreponemal serologic test consist of Wasserman test, venereal disease research laboratory (VDRL), toluidine red unheated serum test (TRUST), unheated serum reagin (USR), rapid plasma reagin (RPR) and treponemal serologic test, such as T. pallidum passive particle agglutination (TPPA), T. pallidum haemagglutination assay (TPHA), fluorescent treponemal antibody absorption (FTA-Abs), enzyme immunoassay (EIA) and rapid test. The algorithm of serologic test can be divided into traditional or reverse.

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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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Diagnosis of Trichomonas vaginalisInfection by PCR Using Vaginal Swab Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.36.11.3205-3210.1998 ◽

1998 ◽

Vol 36 (11) ◽

pp. 3205-3210 ◽

Cited By ~ 119

Author(s):

Guillermo Madico ◽

Thomas C. Quinn ◽

Anne Rompalo ◽

Kelly T. McKee ◽

Charlotte A. Gaydos

Keyword(s):

Trichomonas Vaginalis ◽

Transmitted Disease ◽

Dna Amplification ◽

Vaginal Swab ◽

Specific Method ◽

Analytical Sensitivity ◽

Sexually Transmitted ◽

Tubulin Genes ◽

Joint Strategy ◽

Culture Positive

Trichomonas vaginalis infection is the most prevalent nonviral sexually transmitted disease (STD) in the world. A PCR test using vaginal swab samples for the detection of T. vaginalis was developed to add T. vaginalis infection to the growing list of STDs that can be detected by DNA amplification techniques. A primer set, BTUB 9/2, was designed to target a well-conserved region in the beta-tubulin genes of T. vaginalis. All strains (15 of 15) of T. vaginalistested were successfully detected by PCR giving a single predicted product of 112 bp in gel electrophoresis. No such targeted product was amplified with DNA from Trichomonas tenax,Trichomonas gallinae, Chlamydia trachomatis, Neisseria gonorrhoeae, Giardia lamblia, Chilomastix sulcatus, Dientamoeba fragilis, and Entamoeba histolytica. An optimal analytical sensitivity of one T. vaginalis organism per PCR was achieved. Culture, performed with the Inpouch TV culture system, was examined daily with a light microscope to identify T. vaginalis. Twenty-three of 350 (6.6%) vaginal swab samples from women attending an army medical clinic were culture positive forT. vaginalis. Of these culture positive specimens, PCR detected 22 of 23 (96%) with primer set BTUB 9/2, and wet preparation detected only 12 of 23 (52%). Seventeen specimens were BTUB 9/2-PCR positive and culture negative. Ten of these discordant specimens were determined to be as true positive by PCR using primer sets TVA 5-1/6 and/or AP65 A/B, which target different regions in theT. vaginalis genome, and seven were determined to be false positive. The sensitivity of BTUB 9/2-PCR was 97% and the specificity was 98%. The sensitivities of culture and wet preparation were 70 and 36%, respectively. The diagnosis of T. vaginalis infection by PCR is a sensitive and specific method that could be incorporated into a joint strategy for the screening of multiple STDs by using molecular amplification methods.

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Laboratory Examination of Syphilis

BioScientia Medicina ◽

10.32539/bsm.v5i8.339 ◽

2021 ◽

Vol 5 (8) ◽

pp. 726-745

Author(s):

M. Izazi Hari Purwoko ◽

Mutia Devi ◽

Suroso Adi Nugroho ◽

Fitriani Fitriani ◽

Raden Pamudji ◽

...

Keyword(s):

Venereal Disease ◽

Transmitted Disease ◽

Treponema Pallidum ◽

Clinical Manifestations ◽

Rapid Test ◽

Dark Field ◽

Morphological Observation ◽

Serologic Test ◽

Sexually Transmitted ◽

Antibody Absorption

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Performance Characteristics of a High-Throughput Automated Transcription-Mediated Amplification Test for SARS-CoV-2 Detection

Journal of Clinical Microbiology ◽

10.1128/jcm.01669-20 ◽

2020 ◽

Vol 58 (10) ◽

Cited By ~ 3

Author(s):

Jimmykim Pham ◽

Sarah Meyer ◽

Catherine Nguyen ◽

Analee Williams ◽

Melissa Hunsicker ◽

...

Keyword(s):

Nucleic Acid ◽

High Throughput ◽

Diagnostic Testing ◽

Test System ◽

Nucleic Acid Amplification Test ◽

Performance Characteristics ◽

Nucleic Acid Amplification ◽

Molecular Diagnostic ◽

Specific Method ◽

Analytical Sensitivity

ABSTRACT The COVID-19 pandemic caused by the new SARS-CoV-2 coronavirus has imposed severe challenges on laboratories in their effort to achieve sufficient diagnostic testing capability for identifying infected individuals. In this study, we report the analytical and clinical performance characteristics of a new, high-throughput, fully automated nucleic acid amplification test system for the detection of SARS-CoV-2. The assay utilizes target capture, transcription-mediated amplification, and acridinium ester-labeled probe chemistry on the automated Panther system to directly amplify and detect two separate target sequences in the open reading frame 1ab (ORF1ab) region of the SARS-CoV-2 RNA genome. The probit 95% limit of detection of the assay was determined to be 0.004 50% tissue culture infective dose (TCID50)/ml using inactivated virus and 25 copies/ml (c/ml) using synthetic in vitro transcript RNA targets. Analytical sensitivity (100% detection) was confirmed to be 83 to 194 c/ml using three commercially available SARS-CoV-2 nucleic acid controls. No cross-reactivity or interference was observed with testing of six related human coronaviruses, as well as 24 other viral, fungal, and bacterial pathogens, at high titers. Clinical nasopharyngeal swab specimen testing (n = 140) showed 100%, 98.7%, and 99.3% positive, negative, and overall agreement, respectively, with a validated reverse transcription-PCR nucleic acid amplification test (NAAT) for SARS-CoV-2 RNA. These results provide validation evidence for a sensitive and specific method for pandemic-scale automated molecular diagnostic testing for SARS-CoV-2.

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A Case of Secondary Syphilis with the Extragenital Chancre on the Nipple

Case Reports in Dermatological Medicine ◽

10.1155/2020/2391907 ◽

2020 ◽

Vol 2020 ◽

pp. 1-3

Author(s):

Xiao Yan Wang ◽

Juliandri ◽

Zi Jing Liu ◽

Jia Wen Zhang ◽

Yun Yi Liu ◽

...

Keyword(s):

Skin Biopsy ◽

Sexually Transmitted Disease ◽

Transmitted Disease ◽

Treponema Pallidum ◽

Signs And Symptoms ◽

Secondary Syphilis ◽

Primary Stage ◽

Sexually Transmitted ◽

Serological Examination

Syphilis is a sexually transmitted disease caused by Treponema pallidum. The signs and symptoms of syphilis vary depending on which of the four stages it presents. The primary stage of syphilis classically presents with a painless ulcer (chancre). We report a case of the extragenital chancre on the nipple which is examined from skin biopsy and immunohistochemistry. This case showed that it is important to identify the special site’s pruritus erythema by pathology and serological examination.

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Performance characteristics of a high throughput automated transcription mediated amplification test for SARS-CoV-2 detection

10.1101/2020.07.06.20143719 ◽

2020 ◽

Author(s):

Jimmykim Pham ◽

Sarah Meyer ◽

Catherine Nguyen ◽

Analee Williams ◽

Melissa Hunsicker ◽

...

Keyword(s):

Nucleic Acid ◽

High Throughput ◽

Limit Of Detection ◽

Diagnostic Testing ◽

Test System ◽

Performance Characteristics ◽

Nucleic Acid Amplification ◽

Molecular Diagnostic ◽

Specific Method ◽

Analytical Sensitivity

ABSTRACTThe COVID-19 pandemic caused by the new SARS-CoV-2 coronavirus has imposed severe challenges on laboratories in their effort to achieve sufficient diagnostic testing capability for identifying infected individuals. In this study we report the analytical and clinical performance characteristics of a new, high-throughput, fully automated nucleic acid amplification test system for the detection of SARS-CoV-2. The assay utilizes target capture, transcription mediated amplification, and acridinium ester-labeled probe chemistry on the automated Panther System to directly amplify and detect two separate target sequences in the ORF1ab region of the SARS-CoV-2 RNA genome. The probit 95% limit of detection of the assay was determined to be 0.004 TCID50/ml using inactivated virus, and 25 c/ml using synthetic in vitro transcript RNA targets. Analytical sensitivity (100% detection) was confirmed to be 83 – 194 c/ml using three commercially available SARS-CoV-2 nucleic acid controls. No cross reactivity or interference was observed with testing six related human coronaviruses, as well as 24 other viral, fungal, and bacterial pathogens, at high titer. Clinical nasopharyngeal swab specimen testing (N=140) showed 100%, 98.7%, and 99.3% positive, negative, and overall agreement, respectively, with a validated reverse transcription PCR NAAT for SARS-CoV-2 RNA. These results provide validation evidence for a sensitive and specific method for pandemic-scale automated molecular diagnostic testing for SARS-CoV-2.

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Using Joinpoint Regression for the Analysis of Trends in Syphilis Incidence in Poland in the Years 1950–2017

Studies in Logic, Grammar and Rhetoric ◽

10.2478/slgr-2019-0041 ◽

2019 ◽

Vol 60 (1) ◽

pp. 7-18

Author(s):

Marcin Milewski ◽

Rafał Milewski ◽

Gabriela Sokołowska ◽

Anna Justyna Milewska

Keyword(s):

Transmitted Disease ◽

Treponema Pallidum ◽

Epidemiological Studies ◽

Sexual Contact ◽

Regression Method ◽

Serological Response ◽

Joinpoint Regression ◽

Sexually Transmitted ◽

The Impact ◽

Splitting Time

Abstract Syphilis is a bacterial sexually transmitted disease (STD), whose main route of infection is through sexual contact. In order to diagnose syphilis, Treponema pallidum must be detected in the material sampled from a lesion and a blood test must be performed in order to detect serological response to syphilis. Since 1946, a statutory obligation to report all cases of syphilis has been in force in Poland, which is why data concerning the incidence is available. The aim of this paper is to analyse trends in syphilis incidence in the years 1950–2017 using Joinpoint Regression and to present the impact of prophylaxis and education of society on syphilis prevention. The Joinpoint Regression method indicated the splitting time points of the trend corresponding to real changes in incidence, which corroborates the purpose of using the method in question in epidemiological studies.

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Sonographic appearance of syphilitic induration mimicking squamous cell carcinoma in the lower lip: a case report

Journal of Medical Case Reports ◽

10.1186/s13256-020-02547-x ◽

2020 ◽

Vol 14 (1) ◽

Author(s):

Satomi Asai ◽

Akihiro Kaneko ◽

Tsukumi Matsuda ◽

Noboru Takanashi ◽

Mika Doi ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Color Doppler ◽

Transmitted Disease ◽

Treponema Pallidum ◽

Japanese Woman ◽

Rapid Plasma Reagin ◽

Lower Lip ◽

Sexually Transmitted

Abstract Background Syphilis is a sexually transmitted disease caused by the spirochete Treponema pallidum. Recently, its incidence has been increasing worldwide. We encountered a young woman who presented with induration mimicking squamous cell carcinoma in the lower lip, without major medical conditions. Case presentation A 25-year-old Japanese woman presented with a 1-month history of a painless induration in her lower lip. Because squamous cell carcinoma was suspected, a preoperation work up was performed, including laboratory tests, an ultrasonographic examination, and a biopsy. The ultrasonography findings showed an oval-shaped 17 × 11 × 12 mm tumor-like lesion with heterogeneous internal echo and an indistinct border. A pressure test and color Doppler sonography revealed that the lesion was soft with a very abundant blood flow. These findings suggested the possibility of underlying inflammatory causes rather than a neoplastic tumor. Serology tests for syphilis, including the anti-Treponema pallidum antibody and reactive rapid plasma reagin tests, were positive. The biopsy revealed no malignancy. Finally, she was diagnosed as having primary syphilis and treated with amoxicillin for 28 days. The rapid plasma reagin value gradually decreased and the initial induration in her lower lip disappeared. Conclusion This case highlights the need for prompt examinations for possible underlying infective causes, such as syphilis, when seeing a painless induration with ulcer in the lip. Ultrasonography was helpful in the differential diagnosis of a tumor-like lesion and should be included in addition to syphilis serology tests, such as anti-Treponema pallidum antibody and rapid plasma reagin tests.

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