A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i>
<i>Treponema pallidum</i> subspp. <i>pallidum</i> is a spirochaete bacterium that causes syphilis, one of the most common sexually transmitted diseases. Syphilis progresses through four distinct stages, each characterized by specific symptoms, namely primary, secondary, laten...
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2024-10-01
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| author | Saranthum Phurijaruyangkun Pongbun Tangjitrungrot Pornpun Jaratsing Suphitcha Augkarawaritsawong Khurawan Kumkrong Sawanya Pongparit Pawita Suwanvattana Supatra Areekit Kosum Chansiri Somchai Santiwatanakul |
| author_facet | Saranthum Phurijaruyangkun Pongbun Tangjitrungrot Pornpun Jaratsing Suphitcha Augkarawaritsawong Khurawan Kumkrong Sawanya Pongparit Pawita Suwanvattana Supatra Areekit Kosum Chansiri Somchai Santiwatanakul |
| author_sort | Saranthum Phurijaruyangkun |
| collection | DOAJ |
| description | <i>Treponema pallidum</i> subspp. <i>pallidum</i> is a spirochaete bacterium that causes syphilis, one of the most common sexually transmitted diseases. Syphilis progresses through four distinct stages, each characterized by specific symptoms, namely primary, secondary, latent, and late (tertiary) syphilis. Serology has been considered the primary diagnostic approach. However, it is plagued by problems such as the limited specificity of nontreponemal tests and the inadequate correlation of treponemal tests with disease activity. In this study, we focused on the development of a loop-mediated isothermal amplification assay utilizing hydroxy naphthol blue (LAMP-HNB) for the diagnosis of <i>T. pallidum</i> subspp. <i>pallidum</i>. Specifically, this study seeks to determine the analytical sensitivity (limit of detection; LOD) and analytical specificity. Four hundred clinical serum samples were analyzed for diagnostic sensitivity, specificity, and predictive value, and each technique’s 95% confidence intervals (95% CI, <i>p</i> < 0.05) were evaluated. The limit of detection for polymerase chain reaction with agarose gel electrophoresis (PCR-AGE), the loop-mediated isothermal amplification assay combined with agarose gel electrophoresis (LAMP-AGE), and LAMP-HNB were 116 pg/µL, 11.6 pg/µL, and 11.6 pg/ µL, respectively. Analytical specificity examinations indicated the absence of cross-reactivity with <i>Leptospira interrogans, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa,</i> human immunodeficiency virus (HIV), and healthy human serum in PCR-AGE, LAMP-AGE, and LAMP-HNB. The diagnostic sensitivity, diagnostic specificity, positive predictive value (PPV), and negative predictive value (NPV) for PCR-AGE were 100.00 (100.00)%, 94.50 (94.40–94.60)%, 94.79 (94.69–94.88)%, and 100.00 (100.00)%, respectively. While, for LAMP-AGE and LAMP-HNB, they were 100.00 (100.00)%, 91.00 (90.87–91.13)%, 91.74 (91.63–91.86)%, and 100.00 (100.00)%, respectively. The LAMP-HNB test is simple, rapid, highly sensitive, and highly specific, without requiring expensive equipment. In the future, the LAMP-HNB assay may develop into a single-step diagnostic process, enabling the use as point-of-care testing for the diagnosis, prevention, and management of syphilis infection. |
| format | Article |
| id | doaj-art-2ec51ec61ed444b2a0c7ca17fe813cb9 |
| institution | Kabale University |
| issn | 2076-0817 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
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| series | Pathogens |
| spelling | doaj-art-2ec51ec61ed444b2a0c7ca17fe813cb92024-11-26T18:16:45ZengMDPI AGPathogens2076-08172024-10-01131194910.3390/pathogens13110949A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i>Saranthum Phurijaruyangkun0Pongbun Tangjitrungrot1Pornpun Jaratsing2Suphitcha Augkarawaritsawong3Khurawan Kumkrong4Sawanya Pongparit5Pawita Suwanvattana6Supatra Areekit7Kosum Chansiri8Somchai Santiwatanakul9Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, ThailandCenter of Excellence in Biosensors, Panyananthaphikhu Chonprathan Medical Center, Srinakharinwirot University, Nonthaburi 11120, ThailandCenter of Excellence in Biosensors, Panyananthaphikhu Chonprathan Medical Center, Srinakharinwirot University, Nonthaburi 11120, ThailandFaculty of Medical Technology, Rangsit University, Pathum Thani 12000, ThailandFaculty of Medical Technology, Rangsit University, Pathum Thani 12000, ThailandFaculty of Medical Technology, Rangsit University, Pathum Thani 12000, ThailandMedical Technology and Reference Laboratory for Infectious Diseases, Bamrasnaradura Infectious Diseases Institute, Nonthaburi 11000, ThailandCenter of Excellence in Biosensors, Panyananthaphikhu Chonprathan Medical Center, Srinakharinwirot University, Nonthaburi 11120, ThailandCenter of Excellence in Biosensors, Panyananthaphikhu Chonprathan Medical Center, Srinakharinwirot University, Nonthaburi 11120, ThailandFaculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand<i>Treponema pallidum</i> subspp. <i>pallidum</i> is a spirochaete bacterium that causes syphilis, one of the most common sexually transmitted diseases. Syphilis progresses through four distinct stages, each characterized by specific symptoms, namely primary, secondary, latent, and late (tertiary) syphilis. Serology has been considered the primary diagnostic approach. However, it is plagued by problems such as the limited specificity of nontreponemal tests and the inadequate correlation of treponemal tests with disease activity. In this study, we focused on the development of a loop-mediated isothermal amplification assay utilizing hydroxy naphthol blue (LAMP-HNB) for the diagnosis of <i>T. pallidum</i> subspp. <i>pallidum</i>. Specifically, this study seeks to determine the analytical sensitivity (limit of detection; LOD) and analytical specificity. Four hundred clinical serum samples were analyzed for diagnostic sensitivity, specificity, and predictive value, and each technique’s 95% confidence intervals (95% CI, <i>p</i> < 0.05) were evaluated. The limit of detection for polymerase chain reaction with agarose gel electrophoresis (PCR-AGE), the loop-mediated isothermal amplification assay combined with agarose gel electrophoresis (LAMP-AGE), and LAMP-HNB were 116 pg/µL, 11.6 pg/µL, and 11.6 pg/ µL, respectively. Analytical specificity examinations indicated the absence of cross-reactivity with <i>Leptospira interrogans, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa,</i> human immunodeficiency virus (HIV), and healthy human serum in PCR-AGE, LAMP-AGE, and LAMP-HNB. The diagnostic sensitivity, diagnostic specificity, positive predictive value (PPV), and negative predictive value (NPV) for PCR-AGE were 100.00 (100.00)%, 94.50 (94.40–94.60)%, 94.79 (94.69–94.88)%, and 100.00 (100.00)%, respectively. While, for LAMP-AGE and LAMP-HNB, they were 100.00 (100.00)%, 91.00 (90.87–91.13)%, 91.74 (91.63–91.86)%, and 100.00 (100.00)%, respectively. The LAMP-HNB test is simple, rapid, highly sensitive, and highly specific, without requiring expensive equipment. In the future, the LAMP-HNB assay may develop into a single-step diagnostic process, enabling the use as point-of-care testing for the diagnosis, prevention, and management of syphilis infection.https://www.mdpi.com/2076-0817/13/11/949<i>Treponema pallidum</i>loop-mediated isothermal amplification (LAMP)hydroxy naphthol blue (HNB) |
| spellingShingle | Saranthum Phurijaruyangkun Pongbun Tangjitrungrot Pornpun Jaratsing Suphitcha Augkarawaritsawong Khurawan Kumkrong Sawanya Pongparit Pawita Suwanvattana Supatra Areekit Kosum Chansiri Somchai Santiwatanakul A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> Pathogens <i>Treponema pallidum</i> loop-mediated isothermal amplification (LAMP) hydroxy naphthol blue (HNB) |
| title | A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> |
| title_full | A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> |
| title_fullStr | A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> |
| title_full_unstemmed | A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> |
| title_short | A Loop-Mediated Isothermal Amplification Assay Utilizing Hydroxy Naphthol Blue (LAMP-HNB) for the Detection of <i>Treponema pallidum</i> Subspp. <i>pallidum</i> |
| title_sort | loop mediated isothermal amplification assay utilizing hydroxy naphthol blue lamp hnb for the detection of i treponema pallidum i subspp i pallidum i |
| topic | <i>Treponema pallidum</i> loop-mediated isothermal amplification (LAMP) hydroxy naphthol blue (HNB) |
| url | https://www.mdpi.com/2076-0817/13/11/949 |
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