Use of Fine-Needle Aspiration for Diagnosis of
            <i>Mycobacterium ulcerans</i>
            Infection

Cassisa, Viviane; Chauty, Annick; Marion, Estelle; Ardant, Marie Françoise; Eyangoh, Sara; Cottin, Jane; Aubry, Jacques; Koussemou, Hugues; Lelièvre, Bénédicte; Ferec, Séverine; Tekaia, Fredj; Johnson, Christian; Marsollier, Laurent

doi:10.1128/jcm.00558-10

Cited by 13 publications

(15 citation statements)

References 15 publications

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“…DSE results after Ziehl-Neelsen staining were positive in 47% of qPCR-positive specimens. The levels of sensitivity of DSE were highly dependent on the type of sampling and were 13% in FNA specimens, 52% in swabs, and 82% in skin biopsy specimens (Table 1), which is consistent with the results of a previous study (4).…”

supporting

confidence: 81%

“…M. ulcerans infection is confirmed by laboratory examination, including Ziehl-Neelsen staining, PCR, histology, and/or culture (3). A fine-needle aspiration (FNA) sample may be collected from nonulcerative lesions (nodule, plaque, or edema), and swabs are taken from the undermined edges of ulcerative lesions (4,5). Biopsy specimens can be obtained from a punch biopsy sample or from excised necrotic tissue.…”

mentioning

confidence: 99%

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Establishment of Quantitative PCR (qPCR) and Culture Laboratory Facilities in a Field Hospital in Benin: 1-Year Results

Marion

Ganlonon²,

Claco³

et al. 2014

J Clin Microbiol

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e No simple diagnostic tool is available to confirm Mycobacterium ulcerans infection, which is an emerging disease reported in many rural areas of Africa. Here, we report the 1-year results of a hospital laboratory that was created in an area of endemicity of Benin to facilitate the diagnosis of M. ulcerans infection. Buruli ulcer is a neglected tropical disease occurring mainly in poor, rural communities in West Africa and is caused by the environmental pathogen Mycobacterium ulcerans. This bacillus produces a unique toxin called mycolactone, which has cytotoxic and immunomodulatory activity and often causes severe skin ulcerations, disfigurement, and disability, with children the most affected. A combination of rifampin and streptomycin or rifampin and clarithromycin has been recommended by the World Health Organization (WHO) since 2004 as the first-line treatment (1, 2). M. ulcerans infection is confirmed by laboratory examination, including Ziehl-Neelsen staining, PCR, histology, and/or culture (3). A fine-needle aspiration (FNA) sample may be collected from nonulcerative lesions (nodule, plaque, or edema), and swabs are taken from the undermined edges of ulcerative lesions (4, 5). Biopsy specimens can be obtained from a punch biopsy sample or from excised necrotic tissue. Rapid diagnosis is essential for the management of patients. Since November 2012, the field laboratory established at the Buruli ulcer treatment center in Pobè, Benin, has been fully equipped to carry out M. ulcerans infection diagnosis and has three separate rooms to avoid crosscontamination. Technicians have been trained to perform quantitative PCR (qPCR) and culture in addition to smear examination. Here, we evaluated the 1-year results of this laboratory, which is the first one associated with a specialized treatment center located in a rural area of endemicity and able to carry out qPCR, culture, and direct smear examination (DSE) of a given sample. Detection and quantification of M. ulcerans by qPCR were compared with culture and Ziehl-Neelsen staining results, and the beneficial effects of this laboratory approach on patient management were evaluated.Among 394 specimens analyzed, 203 (51%) were taken by swab, 101 (26%) by FNA, and 90 (23%) by biopsy. The samples were analyzed by molecular tests (qPCR) and microbiological tests (culture and Ziehl-Neelsen staining). Briefly, swabs were rehydrated and biopsy specimens were minced in 2 ml of sterile water. The Kubica method was used to decontaminate 400 l of swab and skin biopsy specimens to isolate M. ulcerans strains (6). FNA specimens were not decontaminated, because the sampling procedure is considered sterile. After inoculation onto Lowenstein-Jensen medium, growth was monitored weekly for 5 months. Ziehl-Neelsen staining was performed as described previously (3). To prepare DNA, 400 l of suspension was centrifuged, resuspended in 50 mM NaOH solution, and heated at 95°C for 10 min. DNA was then purified with a QIAquick PCR purification kit (Qiagen), according to the manufacturer's ...

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confidence: 81%

mentioning

confidence: 99%

Establishment of Quantitative PCR (qPCR) and Culture Laboratory Facilities in a Field Hospital in Benin: 1-Year Results

Marion

Ganlonon²,

Claco³

et al. 2014

J Clin Microbiol

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“…Fine-needle aspirate or swab samples were obtained from patients with suspected Buruli ulcer and sent to Angers University Hospital (Angers, France) for confirmation by quantitative PCR as described ( 6 , 7 ). Of the 283 samples analyzed, 114 (40%) from 108 different patients were PCR positive.…”

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Buruli Ulcer Disease in Republic of the Congo

Marion

Obvala²,

Babonneau³

et al. 2014

Emerg. Infect. Dis.

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“…We also determined equal sensitivities for microscopy of FNA samples (corresponding to 65%, as reported by Eddyani et al [4]) and punch biopsy specimens from nonulcerative lesions. For ulcerative lesions, available data, including our own, suggest that for both diagnostic tests, swabs are clearly superior to tissue samples (2) and that there are no significant differences in sensitivity between the use of FNA samples and that of punch biopsy specimens (4,8).…”

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Efficiency of Fine-Needle Aspiration Compared with Other Sampling Techniques for Laboratory Diagnosis of Buruli Ulcer Disease

et al. 2010

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In accordance with recent WHO recommendations, this study evaluates the sensitivities of PCR and microscopy for fine-needle aspiration (FNA) versus techniques involving swabs and punch biopsy specimens and suggests that FNA can replace punch biopsies for nonulcerative lesions and may serve as an alternative for ulcerative lesions in cases where scarred edges prevent the collection of swabs.Buruli ulcer disease (BUD), caused by Mycobacterium ulcerans, is an emerging disease predominantly affecting West and Central Africa. BUD initially presents as a painless nodule, papule, and plaque (nonulcerative lesions), evolving into a painless ulcer with characteristically undermined edges (ulcerative lesions). Scarring and contractures may cause severe functional disability (9,11,12). Among the currently available diagnostic laboratory methods (microscopy, culture, PCR, and histopathology), PCR provides the highest sensitivity and is therefore regarded as the method of choice for laboratory confirmation. The WHO encourages all countries where BUD is endemic to ensure PCR confirmation of at least 50% of all cases (1,12,13 a The diagnostic results for swabs, fine-needle aspirates (FNA), punch biopsy specimens (PB), and surgically excised tissue (TS) from 110 laboratory-confirmed BUD cases presenting with nonulcerative (n ϭ 37) and ulcerative (n ϭ 73) lesions from Ghana (Agogo Presbyterian Hospital, Agroyesum Hospital, Apromase Hospital, and Dunkwa Governmental Hospital) and Togo (Tsévié Regional Hospital) were analyzed. For subgroups with sample sizes of Ͻ5, the sensitivity was not calculated (NC). NA, not applicable.b The P values comparing the sensitivities of 2 tests for laboratory diagnosis of BUD (PCR and microscopic examination), stratified into 4 techniques of specimen collection (involving swabs, fine-needle aspirates ͓FNA͔, punch biopsy specimens ͓PB͔, and surgically excised tissue ͓TS͔), were as follows: for swabs, Ͻ0.01 for ulcerative lesions; for FNA, Ͻ0.01 for nonulcerative and ulcerative lesions; for PB, Ͻ0.01 for nonulcerative and ulcerative lesions; and for TS, 0.68 for ulcerative lesions. P values of Ͻ0.05 were considered significant.c The P values comparing the sensitivities of 2 out of 4 techniques of specimen collection (involving swabs, fine-needle aspirates ͓FNA͔, punch biopsy specimens ͓PB͔, and surgically excised tissue ͓TS͔), stratified into 2 tests for laboratory diagnosis of BUD (PCR and microscopic examination), are given. P values of Ͻ0.05 were considered significant ‫.)ء(‬ 3732 on August 29, 2018 by guest

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Use of Fine-Needle Aspiration for Diagnosis of Mycobacterium ulcerans Infection

Cited by 13 publications

References 15 publications

Establishment of Quantitative PCR (qPCR) and Culture Laboratory Facilities in a Field Hospital in Benin: 1-Year Results

Establishment of Quantitative PCR (qPCR) and Culture Laboratory Facilities in a Field Hospital in Benin: 1-Year Results

Buruli Ulcer Disease in Republic of the Congo

Efficiency of Fine-Needle Aspiration Compared with Other Sampling Techniques for Laboratory Diagnosis of Buruli Ulcer Disease

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