Rapid and sensitive detection of Chlamydia trachomatis sexually transmitted infections in resource-constrained settings in Thailand at the point-of-care

Somboonna, Naraporn; Choopara, Ilada; Arunrut, Narong; Sukhonpan, Kanchapan; Sayasathid, Jarun; Dean, Deborah; Kiatpathomchai, Wansika

doi:10.1371/journal.pntd.0006900

Cited by 22 publications

(26 citation statements)

References 8 publications

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“…Overall results from 51 positive and negative references as well as real food and clinical samples evaluation demonstrated that our ready-to-use, 1-step ITS1 LAMP-HNB offered high specificity and sensitivity for fungi detection, and the assay was rapid (<1 h) and simple requiring only a water bath (or a heat pad) of 58 • C (Notomi et al, 2000;Somboonna et al, 2018). In addition, this assay gave negative results to common (uncontaminated) food, clinical and environmental (e.g., air, drinking water; Fig.…”

Section: Discussionmentioning

confidence: 93%

“…Thus, using the ITS1 LAMP primers allows later nucleotide sequence probe detections specific for any fungal genera (or species). A specific nucleotide sequence probe, such as in Somboonna et al (2018), may be designed as a second step to selectively identify severely virulent species, for instance. Further, the ability to directly quantitate the color readout into copies of fungi via just a spectrophotometry reading minimizes extra cost and time, and supports local detection where laboratory equipments are limited.…”

Section: Discussionmentioning

confidence: 99%

“…The specificities of the assays were determined with representative fungi (positive) and bacteria (negative) isolates (listed in Table S1). Additional negative controls included crude extracted DNA from human (Homo sapiens) (Somboonna et al, 2018; study was approved by the Institutional Review Board of Buddhachinaraj Phitsanulok Hospital (101/54) and the Ethics Committee for Research in Human Subjects of the Department of Disease Control, Bangkok (FWA00013622)), pork (Sus scrofa), chicken (Gallus gallus) and fish (Pangasius hypophthalmus) meats from local supermarket in Bangkok, Thailand. Crude DNA extraction from additional negative controls was performed by resuspending a sample in 20 µL of Tris-EDTA buffer, boiling at 95 • C for 15 min, and placed on ice prior to centrifugation at 11,772 × g for 10 min.…”

Section: Comparison Of Specificity and Sensitivity (Detection Limit) mentioning

confidence: 99%

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Specific and sensitive, ready-to-use universal fungi detection by visual color using ITS1 loop-mediated isothermal amplification combined hydroxynaphthol blue

Choopara

Teethaisong

Arunrut

et al. 2021

PeerJ

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Being ubiquitous, fungi are common opportunistic pathogens to humans that can lead to invasive and life-threatening infections in immunocompromised individuals. Eukaryote-resembling cell membrane and filamentous branches make the fungal diagnosis difficult. This study therefore developed a ready-to-use ITS1 loop-mediated isothermal amplification combined with hydroxynaphthol blue (LAMP-HNB) for rapid, sensitive and specific colorimetric detection of universal fungi in all phyla. The ITS1 LAMP-HNB could identify every evolutionary phylum of fungi according to sequence analyses. We tested a total of 30 clinically relevant fungal isolates (representing three major human pathogenic phyla of fungi, namely Zygomycota, Ascomycota and Basidiomycota) and 21 non-fungal isolates, and the ITS1 LAMP-HNB properly identified all isolates, with a detection limit of as low as 4.6 ag (9.6 copies), which was identical to ITS1 and 18S rDNA PCR. The assays were also validated on the feasibility of point-of-care diagnostic with real food (dry peanuts, chili and garlics) and blood samples. Furthermore, the shelf life of our ready-to-use ITS1 LAMP activity (≥50%) was more than 40 days at 30 °C with 3–5% polyvinyl alcohol or glycerol additive. The results supported the ready-to-use ITS1 LAMP-HNB for simple detection of fungi contamination with high sensitivity in local and resource-constrained areas to prevent opportunistic fungal species infections.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Comparison Of Specificity and Sensitivity (Detection Limit) mentioning

confidence: 99%

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Specific and sensitive, ready-to-use universal fungi detection by visual color using ITS1 loop-mediated isothermal amplification combined hydroxynaphthol blue

Choopara

Teethaisong

Arunrut

et al. 2021

PeerJ

Self Cite

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“…Substantial research is focused on using isothermal amplification chemistries for diagnosis of infectious disease. For example, chlamydia (caused by the pathogen Chlamydia trachomatis, CT) is the most common sexually transmitted infection worldwide, with more than 110 million cases reported annually (4). Diagnosis of CT infections is challenged by a lack of standard symptoms (many infections are asymptomatic) (5) and the presence of mixed flora (particularly in the female reproductive tract) (6).…”

Section: Introductionmentioning

confidence: 99%

Real-time kinetics and high-resolution melt curves in single-molecule digital LAMP to differentiate and study specific and non-specific amplification

Rolando

Jue

Barlow

et al. 2020

Nucleic Acids Research

150

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Isothermal amplification assays, such as loopmediated isothermal amplification (LAMP), show great utility for the development of rapid diagnostics for infectious diseases because they have high sensitivity, pathogen-specificity and potential for implementation at the point of care. However, elimination of non-specific amplification remains a key challenge for the optimization of LAMP assays. Here, using chlamydia DNA as a clinically relevant target and high-throughput sequencing as an analytical tool, we investigate a potential mechanism of non-specific amplification. We then develop a real-time digital LAMP (dLAMP) with high-resolution melting temperature (HRM) analysis and use this single-molecule approach to analyze approximately 1.2 million amplification events. We show that single-molecule HRM provides insight into specific and non-specific amplification in LAMP that are difficult to deduce from bulk measurements. We use real-time dLAMP with HRM to evaluate differences between polymerase enzymes, the impact of assay parameters (e.g. time, rate or florescence intensity), and the effect background human DNA. By differentiating true and false positives, HRM enables determination of the optimal assay and analysis parameters that leads to the lowest limit of detection (LOD) in a digital isothermal amplification assay.

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“…A LAMP assay developed for the detection of NG showed the same sensitivity of a reference PCR test when used on crude urine [79]. The applicability of using samples without performing traditional DNA extraction was also proven for a LAMP assay used to detect CT in endocervical swabs, with a lysis step of just five minutes before amplification [80].…”

Section: Isothermal Amplification Assays and Microfluidicsmentioning

confidence: 91%

Current and Future Trends in the Laboratory Diagnosis of Sexually Transmitted Infections

Caruso

Giammanco

Virruso

et al. 2021

IJERPH

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Sexually transmitted infections (STIs) continue to exert a considerable public health and social burden globally, particularly for developing countries. Due to the high prevalence of asymptomatic infections and the limitations of symptom-based (syndromic) diagnosis, confirmation of infection using laboratory tools is essential to choose the most appropriate course of treatment and to screen at-risk groups. Numerous laboratory tests and platforms have been developed for gonorrhea, chlamydia, syphilis, trichomoniasis, genital mycoplasmas, herpesviruses, and human papillomavirus. Point-of-care testing is now a possibility, and microfluidic and high-throughput omics technologies promise to revolutionize the diagnosis of STIs. The scope of this paper is to provide an updated overview of the current laboratory diagnostic tools for these infections, highlighting their advantages, limitations, and point-of-care adaptability. The diagnostic applicability of the latest molecular and biochemical approaches is also discussed.

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Rapid and sensitive detection of Chlamydia trachomatis sexually transmitted infections in resource-constrained settings in Thailand at the point-of-care

Cited by 22 publications

References 8 publications

Specific and sensitive, ready-to-use universal fungi detection by visual color using ITS1 loop-mediated isothermal amplification combined hydroxynaphthol blue

Specific and sensitive, ready-to-use universal fungi detection by visual color using ITS1 loop-mediated isothermal amplification combined hydroxynaphthol blue

Real-time kinetics and high-resolution melt curves in single-molecule digital LAMP to differentiate and study specific and non-specific amplification

Current and Future Trends in the Laboratory Diagnosis of Sexually Transmitted Infections

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