Analysis of an IS<i>2404</i>-Based Nested PCR for Diagnosis of Buruli Ulcer Disease in Regions of Ghana Where the Disease Is Endemic

Stienstra, Ymkje; Werf, Tjip S. van der; Guarner, Jeannette; Raghunathan, Pratima L.; Whitney, Ellen A. Spotts; Graaf, Winette T.A. van der; Asamoa, Kwame; Tappero, Jordan W.; Ashford, David A.; King, C. Harold

doi:10.1128/jcm.41.2.794-797.2003

Cited by 38 publications

(32 citation statements)

References 18 publications

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“…Although there was little error by experienced clinicians in endemic areas of ulcers, there were a number of reports of misdiagnosis, which could be as high as 30%. [10][11][12][13][14] The WHO now puts a lot of emphasis on laboratory confirmation, because the introduction of antibiotic therapy results in the formation of a network of laboratories that confirms BU cases. Nevertheless, more emphasis on laboratory support for case management is focusing on IS2404 PCR [15][16][17][18] and sidelining smear microscopy, which is the cornerstone of TB control.…”

Section: Discussionmentioning

confidence: 99%

“…BU cases have been missed initially, 7-9 but presumed BU lesions have proven to be other conditions. [10][11][12][13] In the past, surgery was used as the only means of BU case management; however, the World Health Organization (WHO) currently recommends an 8-week combination therapy with rifampicin and streptomycin as first-line treatment. 14 With the introduction of this antimycobacterial treatment, laboratory confirmation of clinically suspected cases became even more crucial for the clinical management of BU.…”

Section: Introductionmentioning

confidence: 99%

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Combining PCR with Microscopy to Reduce Costs of Laboratory Diagnosis of Buruli Ulcer

Yeboah-Manu

Asante-Poku²,

Asan-Ampah³

et al. 2011

The American Journal of Tropical Medicine and Hygiene

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Combining PCR with Microscopy to Reduce Costs of Laboratory Diagnosis of Buruli Ulcer

Yeboah-Manu

Asante-Poku²,

Asan-Ampah³

et al. 2011

The American Journal of Tropical Medicine and Hygiene

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“…The following three PCR assays were performed as previously described: a single-run gel-based PCR (13), a nested gel-based PCR (11), and a quantitative real-time PCR (7). All PCR assays targeted the insertion sequence IS2404 (8,15).…”

Section: Methodsmentioning

confidence: 99%

Effects of Decontamination, DNA Extraction, and Amplification Procedures on the Molecular Diagnosis of Mycobacterium ulcerans Disease (Buruli Ulcer)

et al. 2012

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We compared two DNA extraction methods (a semiautomated method using a Maxwell kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR, and a real-time quantitative PCR) on 74 surgical tissue specimens from patients with clinically suspected Buruli ulcer. All of these procedures were compared before and after decontamination. We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell 16 extraction method, performed on nondecontaminated suspensions, are the best for the molecular diagnosis of Mycobacterium ulcerans disease. Mycobacterium ulcerans disease, commonly called Buruli ulcer (BU), is a skin disease mainly endemic to certain riverine areas of West and Central Africa (4,12). The disease may present with a diverse range of clinical symptoms, and, due to a possible confusion with other tropical skin diseases, a diagnosis based strictly on clinical observation is not always accurate, leading to the necessity of confirmatory tests such as microscopy, culture, histopathology, and PCR (3, 16).Microscopy is comparatively straightforward to perform, and the materials and skills required are available in regions of endemicity; but diagnosis based on microscopy lacks sensitivity (16). Therefore, even when samples are negative when tested by microscopy, another test should be carried out to confirm the diagnosis. Cultivation of M. ulcerans also lacks sensitivity and takes time to give results (6), rendering it less useful for the routine management of patients. Histopathology is sensitive, specific for BU, and helpful for differential diagnosis but is rarely available in countries where BU is endemic due to a lack of specialists trained and experienced in this technique (16).PCR has been shown to be sensitive as well as specific and is increasingly used for BU diagnosis in countries of endemicity (10,14). Various commercial and in-house DNA extraction and amplification procedures are used, mainly targeting the insertion sequence IS2404 of the M. ulcerans genome. However, only a few studies have evaluated these methods (5). Nevertheless, such an evaluation would be essential to identify the best method for the detection of M. ulcerans by PCR.Since several laboratories which perform PCR also routinely perform culture, PCR is sometimes carried out on suspensions of specimens that have been subjected to a decontamination protocol in preparation for culture. To our knowledge, the effects of this decontamination step on PCR results have not, to date, been investigated.In this study, we compared in two separate laboratories two extraction methods (a semiautomated method using the Maxwell 16 kit [Promega, Leiden, The Netherlands] and a modification of the method of Boom [11]) and three amplification procedures (a single-step PCR, a nested PCR, and a real-time quantitative PCR) routinely performed on surgical tissue specimens from patients with suspected BU. All of these procedures were applied o...

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“…Fixed joint deformities are a consequence of BU particularly when the lesion is close to a joint. 9 For late-presenting lesions, 33 107 In some cases, with advance lesions that have affected joints and bones, amputation may be the only option for these lesions to heal. 108 Lesions in the head and neck region particularly could result in visual impairment and extensive facial scars that have negative impact on the patients including their self-confidence.…”

Section: Disability Stigmatization and Rehabilitationmentioning

confidence: 99%

“…27,28,[31][32][33] With an analytical specificity of 100% and a sensitivity of 79%-98%, IS2404 PCR is considered to be the most reliable technique for the detection of M. ulcerans in human diagnostic samples. These sensitivity and specificity data for PCR are from studies using punch biopsies or swabs.…”

Section: Diagnostic Confirmationmentioning

confidence: 99%

Buruli ulcer: wound care and rehabilitation

Frimpong¹,

Duah²,

Wansbrough‐Jones³

et al. 2016

CWCMR

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Buruli ulcer caused by Mycobacterium ulcerans is a neglected tropical disease characterized by extensive ulceration involving predominantly the upper and lower limbs of patients. The disease is common in rural tropical communities in West and Central Africa, where access to proper health care is limited. Pathogenesis of the characteristic painless ulcers is linked to the elaboration by M. ulcerans of a lipid toxin called mycolactone that has potent cytopathic, immunosuppressive, and analgesic effects on a host of cells in cutaneous tissues. Mycolactone is known to profoundly inhibit secretion of a plethora of proteins that are essential for wound healing. Even though a combination antibacterial therapy of streptomycin and rifampicin for 8 weeks is effective for treatment, it relies on good and appropriate wound management to prevent secondary bacterial infections and improve healing. Evidence-based interventions for wound care in Buruli ulcer disease are often lacking and have relied on expert advice and recommendations. Surgical interventions are limited to debridement of necrotic tissue and grafting of extensive ulcers, usually after antibiotic therapy. Patients' rehabilitation is an important component of care to reduce disabilities associated with the disease and proper integration into the community after treatment.

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Analysis of an IS2404-Based Nested PCR for Diagnosis of Buruli Ulcer Disease in Regions of Ghana Where the Disease Is Endemic

Cited by 38 publications

References 18 publications

Combining PCR with Microscopy to Reduce Costs of Laboratory Diagnosis of Buruli Ulcer

Combining PCR with Microscopy to Reduce Costs of Laboratory Diagnosis of Buruli Ulcer

Effects of Decontamination, DNA Extraction, and Amplification Procedures on the Molecular Diagnosis of Mycobacterium ulcerans Disease (Buruli Ulcer)

Buruli ulcer: wound care and rehabilitation

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