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贝宁野战医院 10 年布路里溃疡分子诊断实验室活动概述。

An Overview of 10 Years of Activity of a Molecular Laboratory for Buruli Ulcer Diagnosis at a Field Hospital in Benin.

机构信息

Universitaire Angers, Nantes Université, CHU Angers, Inserm, INCIT, Angers, France.

Ecole Doctorale en Sciences et Technologie, Université Libanaise, Hadath, Lebanon.

出版信息

J Clin Microbiol. 2023 Jun 20;61(6):e0027423. doi: 10.1128/jcm.00274-23. Epub 2023 May 22.

DOI:10.1128/jcm.00274-23
PMID:37212702
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10281091/
Abstract

Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. Early diagnosis is crucial to prevent morbidity. In November 2012, a field laboratory fully equipped for the rapid on-site quantitative PCR (qPCR) diagnosis of M. ulcerans was established at the Buruli ulcer treatment center (CDTLUB) center in Pobè Benin, a region where BU is endemic. We describe its first 10 years of activity and its gradual evolution into an expert laboratory for BU diagnosis. From 2012 to 2022, the laboratory analyzed 3,018 samples from patients attending consultations for suspected BU at the CDTLUB in Pobè. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. Since 2019, the laboratory has also received and analyzed 570 samples from other centers. The laboratory confirmed the diagnosis of BU by qPCR for 39.7% samples: M. ulcerans DNA was detected in 34.7% of swabs, 47.2% of all fine needle aspiration samples (FNA) and 44.6% of all skin biopsy specimens. Positive Ziehl-Neelsen staining results were obtained for 19.0% samples. Bacterial load, estimated by qPCR, was significantly greater for the Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples, and detection rates were highest for FNA samples. Overall, 26.3% of the samples received from other centers were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado, Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success. Optimal patient care depends on the close proximity of a molecular biology structure to BU treatment centers. Finally, FNA should be promoted among caregivers. Here, we describe the first 10 years of activity at a field laboratory established at the Buruli ulcer treatment center (CDTLUB) in Pobè, Benin, a country in which Mycobacterium ulcerans is endemic. Between 2012 and 2022, the laboratory analyzed 3,018 samples from patients consulting the CDTLUB of Pobè with a suspected clinical BU. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. In total, 39.7% of samples tested positive by qPCR and 19.0% tested positive by Ziehl-Neelsen staining. Detection rates were highest for FNA samples, and the bacterial loads estimated by qPCR were significantly higher for Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples. Since 2019, the laboratory has also analyzed 570 samples received from outside the CDTLUB of Pobè, 26.3% of which were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado in Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success, with major benefits for both the medical staff and patients. Our findings illustrate that the usefulness and feasibility of having a diagnostic center in rural Africa, where the disease is endemic, is a key part of optimal patient care, and that FNA should be promoted to increase detection rates.

摘要

溃疡分枝杆菌病(BU)是一种由溃疡分枝杆菌引起的被忽视的热带病。早期诊断对于预防发病至关重要。2012 年 11 月,在贝宁波贝的溃疡治疗中心(CDTLUB)建立了一个配备齐全的现场实验室,用于快速现场定量 PCR(qPCR)诊断溃疡分枝杆菌病,该地区是 BU 的流行地区。我们描述了其第一个 10 年的活动及其逐渐演变为 BU 诊断的专家实验室。从 2012 年到 2022 年,该实验室分析了来自波贝 CDTLUB 就诊的疑似 BU 患者的 3018 个样本。进行了齐尔-尼尔森染色和针对 IS2404 序列的 qPCR。自 2019 年以来,该实验室还接收和分析了来自其他中心的 570 个样本。实验室通过 qPCR 确认了 BU 的诊断:在 39.7%的样本中检测到溃疡分枝杆菌 DNA:在 34.7%的拭子、47.2%的所有细针抽吸样本(FNA)和 44.6%的所有皮肤活检标本中检测到 M. ulcerans DNA。19.0%的样本得到了阳性齐尔-尼尔森染色结果。通过 qPCR 估计的细菌负荷在齐尔-尼尔森阳性样本中显著大于齐尔-尼尔森阴性样本,并且 FNA 样本的检测率最高。总体而言,来自其他中心的 26.3%的样本对 BU 呈阳性。这些样本大多来自贝宁的拉洛、阿拉达和扎格纳多的 CDTLUB。在波贝的 CDTLUB 建立实验室取得了巨大的成功。最佳的患者护理取决于分子生物学结构与 BU 治疗中心的紧密接近。最后,应该推广在护理人员中进行 FNA。在这里,我们描述了贝宁波贝溃疡治疗中心(CDTLUB)建立的现场实验室的前 10 年活动,该实验室在溃疡分枝杆菌流行的国家建立。2012 年至 2022 年间,该实验室分析了来自波贝 CDTLUB 就诊的疑似临床 BU 患者的 3018 个样本。进行了齐尔-尼尔森染色和针对 IS2404 序列的 qPCR。共有 39.7%的样本通过 qPCR 检测为阳性,19.0%的样本通过齐尔-尼尔森染色检测为阳性。FNA 样本的检测率最高,通过 qPCR 估计的细菌负荷在齐尔-尼尔森阳性样本中显著高于齐尔-尼尔森阴性样本。自 2019 年以来,该实验室还分析了来自波贝 CDTLUB 以外的 570 个样本,其中 26.3%的样本对 BU 呈阳性。这些样本大多来自贝宁的拉洛、阿拉达和扎格纳多的 CDTLUB。在波贝的 CDTLUB 建立实验室取得了巨大的成功,为医务人员和患者都带来了重大的利益。我们的研究结果表明,在疾病流行的非洲农村地区建立一个诊断中心对于最佳患者护理是有用且可行的,并且应该推广 FNA 以提高检测率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/3085d1a0eba8/jcm.00274-23-f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/f0fb6c441bb3/jcm.00274-23-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/38f0bbdbd643/jcm.00274-23-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/2ce17983cd8a/jcm.00274-23-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/3085d1a0eba8/jcm.00274-23-f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/f0fb6c441bb3/jcm.00274-23-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/38f0bbdbd643/jcm.00274-23-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/2ce17983cd8a/jcm.00274-23-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea76/10281091/3085d1a0eba8/jcm.00274-23-f004.jpg

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