Rapid and sensitive detection of Chlamydia trachomatis sexually transmitted infections in resource-constrained settings in Thailand at the point-of-care
| Autor: | Deborah Dean, Kanchapan Sukhonpan, Naraporn Somboonna, Wansika Kiatpathomchai, Narong Arunrut, Ilada Choopara, Jarun Sayasathid |
|---|---|
| Přispěvatelé: | Small, Pamela LC |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Male Time Factors RC955-962 Prevalence Artificial Gene Amplification and Extension Chlamydia trachomatis medicine.disease_cause Pathology and Laboratory Medicine Medical and Health Sciences Polymerase Chain Reaction law.invention From Innovation to Application Spectrum Analysis Techniques law Arctic medicine. Tropical medicine Medicine and Health Sciences Medicine Chlamydia DNA extraction Polymerase chain reaction Thermal cycler Amplicon Biological Sciences Thailand Bacterial Pathogens Bioassays and Physiological Analysis Infectious Diseases Medical Microbiology Spectrophotometry Viral Pathogens Viruses Female Public aspects of medicine RA1-1270 Pathogens Papillomaviruses Point-of-Care Systems 030106 microbiology Loop-mediated isothermal amplification Sexually Transmitted Diseases Research and Analysis Methods Microbiology Sensitivity and Specificity 03 medical and health sciences Extraction techniques Tropical Medicine Nucleic Acid Amplification Tests Humans Molecular Biology Techniques Microbial Pathogens Molecular Biology Colorimetric Assays Bacteria business.industry Public Health Environmental and Occupational Health Organisms Biology and Life Sciences Human Papillomavirus Chlamydia Infections Virology 030104 developmental biology Good Health and Well Being Ultraviolet-Visible Spectroscopy business DNA viruses Biochemical Analysis |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS neglected tropical diseases, vol 12, iss 12 PLoS Neglected Tropical Diseases, Vol 12, Iss 12, p e0006900 (2018) |
| ISSN: | 1935-2735 1935-2727 |
| Popis: | Chlamydia trachomatis is the leading cause of sexually transmitted diseases (STDs) in females and males in both developed and developing countries, with more than 110 million cases annually. C. trachomatis resists antibiotic treatment and is a cofactor in HIV transmission and human cervical cancer [1]. Infection is often asymptomatic, causing the epidemiology to be underestimated. Therefore, we have developed a rapid, inexpensive, easy-to-interpret, sensitive and specific point-of-care (POC) C. trachomatis detection system, using loop-mediated isothermal amplification (LAMP) for target C. trachomatis DNA amplification, followed by gold nanoparticle probe (AuNP) for colorimetric C. trachomatis specific readout. The assay was evaluated using clinical samples and compared with polymerase chain reaction (PCR) of the same target gene, which is an outer membrane protein A (ompA) gene and a respected standard for C. trachomatis detection [1,2]. For nucleic acid amplification tests, recently LAMP has presented an attractive alternative to standard methods like PCR due to its low price, ease of use, rapid results, and lack of requirement for an expensive thermal cycler and specialized kits for DNA extraction and purification. A simple 5-minute boil for crude DNA lysis is sufficient for the LAMP reaction because the Bacillus stearothermophilus (Bst) DNA polymerase in LAMP has fewer inhibitor problems than Thermus aquaticus (Taq) DNA polymerase in PCR [3]. LAMP amplifies the target at a single temperature with high sensitivity (from 10 to 100 genome copies), and the product can be visualized as a white magnesium pyrophosphate precipitate. However, nebulous precipitate has the potential to cause misreading [3,4], so analysis by agarose gel-electrophoresis (GE), as with PCR, is standard. The positive LAMP reaction appears as multiple bands because several primers amplify amplicons of different sizes that are intercalated. Nevertheless, GE requires an electrophoresis apparatus, time, and often ethidium bromide exposure. In the current study, crude DNA lysis with combined LAMP-AuNP for POC C. trachomatis detection has been developed, and the specificity and limit of detection were validated by several positive and negative C. trachomatis genomes, as well as the possibility for POC diagnostic based on a statistical number of clinical endocervical swab sample tests (see Box 1). Box 1: Advantages and disadvantages of C. trachomatis LAMP-AuNP. Advantages Sensitive to as low as 11.25 copies of target DNA Total assay time is |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |