Rapid molecular diagnosis of chronic skin ulcers

Pahil, Sapna; Goyal, Kapil

doi:10.1016/s2214-109x(14)70242-6

Cited by 3 publications

(5 citation statements)

References 7 publications

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“…Operating this technique at the point of care is very challenging. This is because of its requirement for sophisticated laboratories, equipped with expensive equipment such as a thermocycler [12], which can cost more than 15 times as much as isothermal fluorometers required for the performance RPA. RPA performance requires less costly equipment and can be performed in less resource settings in a mobile suitcase laboratory [18,20].…”

Section: Discussionmentioning

confidence: 99%

“…However, PCR techniques can only be applied in a reference laboratory and not at the point of care (POC) in endemic communities. As such, the need for diagnostic techniques that do not involve costly equipment and can be used in remote or resource limited areas have been indicated [12]. The use of isothermal amplification techniques such as Loop-mediated amplification (LAMP), for on-site screening has been developed as a possible POC test for chronic skin ulcers caused by T. pallidum and H. ducreyi [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

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Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions

et al. 2020

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Yaws is a skin debilitating disease caused by Treponema pallidum subspecies pertenue with most cases reported in children. World Health Organization (WHO) aims at total eradication of this disease through mass treatment of suspected cases followed by an intensive follow-up program. However, effective diagnosis is pivotal in the successful implementation of this control program. Recombinase polymerase amplification (RPA), an isothermal nucleic acid amplification technique offers a wider range of differentiation of pathogens including those isolated from chronic skin ulcers with similar characteristics such as Haemophilus ducreyi (H. ducreyi). We have developed a RPA assay for the simultaneous detection of Treponema pallidum (T. pallidum) and H. ducreyi (TPHD-RPA). The assay demonstrated no cross-reaction with other pathogens and enable detection of T. pallidum and H. ducreyi within 15 min at 42 °C. The RPA assay was validated with 49 clinical samples from individuals confirmed to have yaws by serological tests. Comparing the developed assay with commercial multiplex real-time PCR, the assay demonstrated 94% and 95% sensitivity for T. pallidum and H. ducreyi, respectively and 100% specificity. This simple novel TPHD-RPA assay enables the rapid detection of both T. pallidum and H. ducreyi in yaws-like lesions. This test could support the yaws eradication efforts by ensuring reliable diagnosis, to enable monitoring of program success and planning of follow-up interventions at the community level.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions

et al. 2020

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“…However, this technique can only be applied in a reference laboratory and not at the point of care in endemic communities. As such, the need for diagnostic techniques that do not involve costly equipment and can be used in remote areas have been indicated [12]. The use of isothermal amplification techniques such as Loop-mediated amplification (LAMP) for on-site screening has been proposed as a future POC test for chronic skin ulcers caused by T. pallidum and H. ducreyi [13,14].…”

Section: Nucleic Acid Amplification Technique Especially Polymerase Cmentioning

confidence: 99%

“…ducreyi lesions [7,8,11]. However, operating this technique at the point of care is very challenging because of its requirement for sophisticated laboratories equipped with expensive equipment such as a thermocycler [12] which can cost more than 15 times as much as isothermal fluorometers which enable the performance RPA. RPA performance requires less costly equipment and can be performed in less resource settings in a mobile suitcase laboratory.…”

Section: Analytical Sensitivity and Specificity Of Duplex Tphd Rpa Assaymentioning

confidence: 99%

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in yaws-like lesions

Frimpong¹,

Simpson²,

Ahor³

et al. 2020

Preprint

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Yaws is a skin debilitating disease caused by Treponema pallidum subspecies pertenue with most cases reported in children. World Health Organization (WHO) aims at total eradication of this disease through mass treatment of suspected cases followed by an intensive follow-up program. However, effective diagnosis is pivotal in the successful implementation of this control program. Recombinase polymerase amplification (RPA), an isothermal nucleic acid amplification technique offers a wider range of differentiation of pathogens including those isolated from chronic skin ulcers with similar characteristics such as Haemophilus ducreyi (H. ducreyi). We have developed a duplex RPA assay for the simultaneous detection of Treponema pallidum (T. pallidum) and H. ducreyi (TPHD-RPA). TPHD-RPA assay demonstrated no cross-reaction with other pathogens and enable detection of T. pallidum and H. ducreyi within 15 minutes at 42 oC. The duplex RPA assay was validated with 49 clinical samples from individuals confirmed to have yaws by serological tests. Compared with commercial multiplex real-time PCR, the TPHD-RPA assay demonstrated 94-95% sensitivity for T. pallidum and H. ducreyi confirmed samples, respectively and 100% specificity. This simple novel TPHD-RPA assay enables the rapid detection of both T. pallidum and H. ducreyi in yaws-like lesions. This test could support the yaws eradication programs by ensuring effective diagnosis as well as enable monitoring of eradication efforts success or failure and planning of follow-up interventions at the community level.

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“…Post-eradication surveillance following the currently ongoing mass-azithromycin treatment phase [ 4 ] would benefit from rapid and cost-effective molecular tests that are able to distinguish TPE infection [ 18 ] from infections with all other TP subspecies ( TPA and TEN ) and bacteria that are involved in tropical skin ulcers and which may fall together with TP seropositivity. Potentially a single overlooked yaws case would result in a failure of global yaws eradication.…”

Section: Introductionmentioning

confidence: 99%

Gene target selection for loop-mediated isothermal amplification for rapid discrimination of Treponema pallidum subspecies

et al. 2018

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We show proof of concept for gene targets (polA, tprL, and TP_0619) that can be used in loop-mediated isothermal amplification (LAMP) assays to rapidly differentiate infection with any of the three Treponema pallidum subspecies (pallidum (TPA), pertenue (TPE), and endemicum (TEN)) and which are known to infect humans and nonhuman primates (NHPs). Four TPA, six human, and two NHP TPE strains, as well as two human TEN strains were used to establish and validate the LAMP assays. All three LAMP assays were highly specific for the target DNA. Amplification was rapid (5–15 min) and within a range of 10E+6 to 10E+2 of target DNA molecules. Performance in NHP clinical samples was similar to the one seen in human TPE strains. The newly designed LAMP assays provide proof of concept for a diagnostic tool that enhances yaws clinical diagnosis. It is highly specific for the target DNA and does not require expensive laboratory equipment. Test results can potentially be interpreted with the naked eye, which makes it suitable for the use in remote clinical settings.

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Rapid molecular diagnosis of chronic skin ulcers

Cited by 3 publications

References 7 publications

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in yaws-like lesions

Gene target selection for loop-mediated isothermal amplification for rapid discrimination of Treponema pallidum subspecies

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