A molecular screening assay to identify <i>Chlamydia trachomati</i>s and distinguish new variants of <i>C. trachomatis</i> from wild‐type

Xiu, Leshan; Li, Yamei; Zhang, Chi; Li, Yizhun; Zeng, Yaling; Wang, Feng; Peng, Junping

doi:10.1111/1751-7915.13724

Cited by 7 publications

(6 citation statements)

References 31 publications

(64 reference statements)

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“…Recent taxonomic developments based on 16S and 23S rRNA gene sequences, along with the development of molecular tests, have divided the Chlamydiaceae family into two genera and nine species, including 5 species found to infect humans [6]. Over time, new variants of C. trachomatis have emerged, and new test techniques that are inexpensive and easily applicable may emerge to identify them [7]. Chlamydia is the most commonly diagnosed bacterium among sexually transmitted diseases [8].…”

Section: Introductionmentioning

confidence: 99%

Molecular Approaches to the Diagnosis of Chlamydia

Konuk¹

2023

Infectious Diseases

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Chlamydia trachomatis is known as the most common bacterial infection agent to pass with sexual transition. This microorganism is an obligatory intracellular parasite. A variety of infections are caused by C. trachomatis, including trachoma, pneumonias in newborns, genital and urinary tract infections, and lymphogranuloma venereum (LGV), which is caused by LGV strains. The diagnosis of Chlamydia trachomatis can be made by cultures and isolations, antigens and antibodies (direct fluorescence, enzyme immunoassays), hybridization, or polymerase chain reaction (PCR). Each year, infection and diagnosis rates increase in the developed world. Since Chlamydia is mostly asymptomatic, screening, and treatment are a key to detecting cases. Polymerase chain reaction (PCR), ligase chain reaction (LCR), and nucleic acid sequence-based amplification (NASBAa) molecular methods can be used for the detection, low concentration, quantification, and identification of organisms. While the traditional PCR method confirms its existence, it can quantify real-time PCR (RT-PCR). This method (RT-PCR) may have low sensitivity among variants of the same species. Also, PCR scans, which receive urine service, offer great advantages. PCR from initial void urine (FVU) samples is highly sensitive in detecting the organism. Urine Chlamydia screenings are more acceptable in large populations and asymptomatic detections.

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Section: Introductionmentioning

confidence: 99%

Molecular Approaches to the Diagnosis of Chlamydia

Konuk¹

2023

Infectious Diseases

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“…In this study, we propose a methodology based on high‐resolution melting (HRM) analysis for SARS‐CoV‐2 detection that can rapidly and consistently identify the virus, subtypes and mutations without requiring a sequencing analysis step, thereby avoiding the need for reagents and/or supplies. HRM is a convenient, extensible, reliable, closed‐tube and cost‐effective approach that has been used for identification of species or mutation scanning or molecular typing in several research fields, such as epidemiology and microbiology (Montgomery et al ., 2010 ; Tong and Giffard, 2012 ; Li et al ., 2014 ; Xiu et al ., 2020b ; Xiu et al ., 2020 ). By exploring the advantages of HRM analysis, we developed a multiplex assay using reverse transcription coupled with HRM analysis of the amplification products to rapidly detect viral RNA and simultaneously identify mutations in SARS‐CoV‐2.…”

Section: Introductionmentioning

confidence: 99%

Detection and classification of SARS‐CoV‐2 using high‐resolution melting analysis

Sun

Xiu

Zhang

et al. 2022

Microbial Biotechnology

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Summary Coronavirus disease 2019 (COVID‐19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS ‐ CoV ‐ 2), has recently posed a significant threat to global public health. The objective of this study was to develop and evaluate a rapid, expandable and sequencing‐free high‐resolution melting (HRM) approach for the direct detection and classification of SARS‐CoV‐2. Thirty‐one common pathogens that can cause respiratory tract infections were used to evaluate the specificity of the method. Synthetic RNA with serial dilutions was utilized to determine the sensitivity of the method. Finally, the clinical performance of the method was assessed using 290 clinical samples. The one‐step multiplex HRM could accurately identify SARS‐CoV‐2 and differentiate mutations in each marker site within approximately 2 h. For each target, the limit of detection was lower than 10 copies/reaction, and no cross‐reactivity was observed among organisms within the specificity testing panel. The method showed good uniformity for SARS‐CoV‐2 detection with a consistency of 100%. Regarding the clade classification performance, the results showed good concordance compared with sequencing, with the rate of agreement being 95.1% (78/82). The one‐step multiplex HRM method is a rapid method for SARS‐CoV‐2 detection and classification.

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“…This novel method is based on a high-resolution melting (HRM) analysis with unlabeled probes and complements current M. genitalium detection using RT-PCR and WGS. HRM is a convenient, closed-tube, and cost-efficient method that is widely used in several research fields, including variant scanning, species identification, and molecular typing ( 19 – 22 ). Although HRM is superior to RT-PCR for identifying many small insertions or deletions and complex mutations, differentiating between two or more possible single nucleotide polymorphisms at a site can be problematic when probes are not used ( 23 , 24 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Antimicrobial Resistance in Mycoplasma genitalium by High-Resolution Melting Analysis with Unlabeled Probes

Xiu

Wang

et al. 2022

Microbiol Spectr

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Mycoplasma genitalium was recently added to the antimicrobial-resistant (AMR) threats “watch list” of the U.S. Centers for Disease Control and Prevention because this pathogen has become extremely difficult to treat as a result of increased resistance. M. genitalium is also difficult to culture, and therefore, molecule detection is the only method available for AMR testing.

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A molecular screening assay to identify Chlamydia trachomatis and distinguish new variants of C. trachomatis from wild‐type

Cited by 7 publications

References 31 publications

Molecular Approaches to the Diagnosis of Chlamydia

Molecular Approaches to the Diagnosis of Chlamydia

Detection and classification of SARS‐CoV‐2 using high‐resolution melting analysis

Rapid Detection of Antimicrobial Resistance in Mycoplasma genitalium by High-Resolution Melting Analysis with Unlabeled Probes

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