Implementation of a National Reference Laboratory for Buruli Ulcer Disease in Togo

Abstract

BackgroundIn a previous study PCR analysis of clinical samples from suspected cases of Buruli ulcer disease (BUD) from Togo and external quality assurance (EQA) for local microscopy were conducted at an external reference laboratory in Germany. The relatively poor performance of local microscopy as well as effort and time associated with shipment of PCR samples necessitated the implementation of stringent EQA measures and availability of local laboratory capacity. This study describes the approach to implementation of a national BUD reference laboratory in Togo.MethodologyLarge scale outreach activities accompanied by regular training programs for health care professionals were conducted in the regions “Maritime” and “Central,” standard operating procedures defined all processes in participating laboratories (regional, national and external reference laboratories) as well as the interaction between laboratories and partners in the field. Microscopy was conducted at regional level and slides were subjected to EQA at national and external reference laboratories. For PCR analysis, sample pairs were collected and subjected to a dry-reagent-based IS2404-PCR (DRB-PCR) at national level and standard IS2404 PCR followed by IS2404 qPCR analysis of negative samples at the external reference laboratory.Principal FindingsThe inter-laboratory concordance rates for microscopy ranged from 89% to 94%; overall, microscopy confirmed 50% of all suspected BUD cases. The inter-laboratory concordance rate for PCR was 96% with an overall PCR case confirmation rate of 78%. Compared to a previous study, the rate of BUD patients with non-ulcerative lesions increased from 37% to 50%, the mean duration of disease before clinical diagnosis decreased significantly from 182.6 to 82.1 days among patients with ulcerative lesions, and the percentage of category III lesions decreased from 30.3% to 19.2%.ConclusionsHigh inter-laboratory concordance rates as well as case confirmation rates of 50% (microscopy), 71% (PCR at national level), and 78% (including qPCR confirmation at external reference laboratory) suggest high standards of BUD diagnostics. The increase of non-ulcerative lesions, as well as the decrease in diagnostic delay and category III lesions, prove the effect of comprehensive EQA and training measures involving also procedures outside the laboratory.

Effectiveness of Routine BCG Vaccination on Buruli Ulcer Disease: A Case-Control Study in the Democratic Republic of Congo, Ghana and Togo

“…This collaboration is supported by the Togolese National Buruli Ulcer Control Program (“Programme National de Lutte contre L'Ulcère de Buruli – Lèpre et Pian” [PNLUB-LP]), in the area of training, active case finding, laboratory confirmation, and treatment of BUD. In 2007, the CHR Maritime was appointed National Reference Centre for BUD in Togo [42] , [43] .…”

“…Data collection was conducted by means of the WHO “BU01” form, and standardized project-specific “BuruliVac” laboratory data entry forms (Form S1). All socio-demographic, clinical, and laboratory data were entered in a web-based database specifically designed for the “BuruliVac” project [43] . Following WHO guidance, the categories of BUD were defined as follows: Category I were single lesions <5 cm in diameter; Category II were single lesions between 5 and 15 cm in diameter; Category III were single lesions >15 cm in diameter, multiple lesions, lesions at critical sites or osteomyelitis [44] .…”

“…In Ghana and Togo, diagnostic samples were collected according to standardized procedures [43] . Briefly, swabs were collected by circling the entire undermined edges of ulcerative lesions.…”

Comparison of Two Assays for Molecular Determination of Rifampin Resistance in Clinical Samples from Patients with Buruli Ulcer Disease

“…To determine the performances of the two assays on clinical specimens, 133 whole-genome extracts from IS2404-positive samples collected before the onset of treatment (swab samples, n ϭ 63; fine-needle aspirates [FNA], n ϭ 40; 3-mm punch biopsy specimens, n ϭ 30) from 91 BUD patients from Togo (17,18) were assessed. P values of Ͻ0.05 were considered significant.…”

First isolation of Mycobacterium ulcerans from Swabs and Fine-Needle-Aspiration Specimens in Togo.

“…In Togo, studies have been done on the risk factors for Mycobacterium ulcerans infection and the detection of cases by the Ziehl-Neelsen and PCR technique on clinical and environmental samples [2,16,17], and to date no data of isolates from clinical samples are available. The objective of this study is to perform an in vitro culture method to isolate circulating clinical strains of M. ulcerans in Togo from fine needle aspiration (FNA) and swabs samples.…”