Development a rapid and accurate multiplex real time PCR method for the detection <i>Chlamydia trachomatis</i> and <i>Mycoplasma hominis</i>

Safarkar, Roya; Mehrabadi, Jalil Fallah; Noormohammadi, Zahra; Mirnejad, Reza

doi:10.1002/jcla.22126

Cited by 10 publications

(8 citation statements)

References 28 publications

(75 reference statements)

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“…Compared with cell cultivation, enzyme immunoassay and direct fluorescence assay are relatively simple and rapid, but are not recommended as routine diagnostic methods due to low diagnostic accuracy (Peng et al, 2020 ). Nucleic acid amplification tests are more sensitive and have an excellent specificity, which is recommended as the new gold diagnostic method for C. trachomatis detection (Herbst De Cortina et al, 2016 ; Safarkar et al, 2017 ). However, their use is confined due to requiring complicated laboratory facilities and trained technicians in resource-impoverished regions.…”

Section: Discussionmentioning

confidence: 99%

“…Hence, it is rarely performed in routine clinical diagnostic application. Nucleic acid amplification technologies (NAATs), including polymerase chain reaction (PCR) and ligase chain reaction, have been considered as a major breakthrough and have been regarded as the new gold standard for the detection of C. trachomatis infection for high sensitivity, specificity, and automation (Gaydos et al, 2013 ; Safarkar et al, 2017 ). Nevertheless, NAATs are often unaffordable and inaccessible in resource-limited regions due to requiring skilled technicians and expensive analytical instruments.…”

Section: Introductionmentioning

confidence: 99%

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Nanoparticle-Based Lateral Flow Biosensor Integrated With Loop-Mediated Isothermal Amplification for Rapid and Visual Identification of Chlamydia trachomatis for Point-of-Care Use

Chen

Zhou

Tan³

et al. 2022

Front. Microbiol.

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Chlamydial infection, caused by Chlamydia trachomatis, is the most common bacterial sexually transmitted infection and remains a major public health problem worldwide, particularly in underdeveloped regions. Developing a rapid and sensitive point-of-care (POC) testing for accurate screening of C. trachomatis infection is critical for earlier treatment to prevent transmission. In this study, a novel diagnostic assay, loop-mediated isothermal amplification integrated with gold nanoparticle-based lateral flow biosensor (LAMP-LFB), was devised and applied for diagnosis of C. trachomatis in clinical samples. A set of LAMP primers based on the ompA gene from 14 C. trachomatis serological variants (serovar A-K, L1, L2, L3) was successfully designed and used for the development of C. trachomatis-LAMP-LFB assay. The optimal reaction system can be performed at a constant temperature of 67°C for 35 min. The total assay process, including genomic DNA extraction (~15 min), LAMP reaction (35 min), and LFB readout (~2 min), could be finished within 60 min. The C. trachomatis-LAMP-LFB could detect down to 50 copies/ml, and the specificity was 100%, no cross-reactions with other pathogens were observed. Hence, our C. trachomatis-LAMP-LFB was a rapid, reliable, sensitive, cost-effective, and easy-to-operate assay, which could offer an attractive POC testing tool for chlamydial infection screening, especially in resource starvation settings.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Nanoparticle-Based Lateral Flow Biosensor Integrated With Loop-Mediated Isothermal Amplification for Rapid and Visual Identification of Chlamydia trachomatis for Point-of-Care Use

Chen

Zhou

Tan³

et al. 2022

Front. Microbiol.

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“…The Mycoplasma species is extremely small and lacks a peptidoglycan cell wall; therefore, it does not appear on gram stain, making it particularly difficult to identify using traditional techniques. The most reliable test for detection of M. hominis is the 16S ribosomal RNA gene by PCR [11], although multiplex real time PCR tests are also being developed [12]. However, these tests are not available at all institutions and often takes days to weeks to obtain results.…”

Section: Discussionmentioning

confidence: 99%

Primary Peritonitis: An Index Case of Mycoplasma hominis Infection in a Healthy Female

Drexel

Tseng

2018

Case Reports in Surgery

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Introduction Primary peritonitis in healthy immunocompetent individuals is rare. Several case reports of Streptococcus species causing peritonitis have been described. Here, we present the first case of Mycoplasma hominis as the cause of primary peritonitis in a healthy woman. Case Report A 42-year-old female with history of uterine fibroids was admitted with abdominal pain and intraperitoneal fluid of unknown etiology. She was initially managed nonoperatively and empirically treated with broad spectrum antibiotics. Blood and urine cultures were unrevealing. Increasing abdominal pain and peritoneal fluid prompted diagnostic laparoscopy which revealed a dense fibrinous exudate covering the entire peritoneal cavity. Peritoneal fluid and biopsies were sent for cytology and culture. The peritoneal fluid was eventually sent for 16 s ribosomal analysis, which discovered Mycoplasma hominis RNA. Her antibiotics were narrowed, and she eventually made a full recovery. Discussion M. hominis is a rare source of systemic infection but has been known to colonize the urogenital tract and cause localized infections. This is the first presentation of M. hominis causing primary peritonitis in a healthy immunocompetent female. Multidisciplinary management of these patients is critical to achieve a timely diagnosis. Surgical exploration is often unavoidable to rule out secondary peritonitis.

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“…Compared with traditional PCR, RT-PCR has several advantages such as high speed, sensitivity, and specificity, and no post-PCR contamination 6) . In our case, we could confirm the results by RT- although their safety in newborns has not been clearly established 8,9) .…”

Section: Case Reportmentioning

confidence: 99%

Successful Treatment of Mycoplasma hominis Meningitis, Diagnosed Using Real-Time Polymerase Chain Reaction, with Ciprofloxacin in a Neonate

Gwon¹,

Song²,

Sohn³

et al. 2020

Neonatal Med

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Mycoplasma hominis can cause life-threatening central nervous system infections in neonates following intrauterine infection or during delivery. In newborns, the diagnosis and treatment of M. hominis meningitis are challenging, because cultures are often negative and the bacterium is not susceptible to empirical antibiotics. Herein, we describe a case of neonatal M. hominis meningitis diagnosed using real-time polymerase chain reaction (RT-PCR) and treated with ciprofloxacin. The patient was a 3-day-old female hospitalized for a fever and lethargy. Her blood laboratory findings were non-specific; cerebrospinal fluid (CSF) examination showed a white blood cell count of 580/µL and indicated meningitis. Her symptoms could not be controlled with empirical antibiotics. Urine culture on a special medium revealed ciprofloxacinsusceptible M. hominis. Furthermore, the RT-PCR performed with the CSF sample revealed M. hominis. Therefore, the patient was administered cipro floxacin; after 2 days, the fever subsided. The patient was discharged on day 30 without complications.

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Development a rapid and accurate multiplex real time PCR method for the detection Chlamydia trachomatis and Mycoplasma hominis

Cited by 10 publications

References 28 publications

Nanoparticle-Based Lateral Flow Biosensor Integrated With Loop-Mediated Isothermal Amplification for Rapid and Visual Identification of Chlamydia trachomatis for Point-of-Care Use

Nanoparticle-Based Lateral Flow Biosensor Integrated With Loop-Mediated Isothermal Amplification for Rapid and Visual Identification of Chlamydia trachomatis for Point-of-Care Use

Primary Peritonitis: An Index Case of Mycoplasma hominis Infection in a Healthy Female

Successful Treatment of Mycoplasma hominis Meningitis, Diagnosed Using Real-Time Polymerase Chain Reaction, with Ciprofloxacin in a Neonate

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