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比较显微镜检查和 Xpert MTB 诊断疑似结核分枝杆菌感染:开普敦教学医院常规诊断的回顾性分析。

Comparison of microscopic and xpert MTB diagnoses of presumptive mycobacteria tuberculosis infection: retrospective analysis of routine diagnosis at Cape Coast Teaching Hospital.

机构信息

Biomedical and Clinical Research Centre, College of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana.

Department of Biomedical Sciences, School of Allied Health Sciences, College of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana.

出版信息

BMC Infect Dis. 2024 Jul 2;24(1):660. doi: 10.1186/s12879-024-09566-9.

Abstract

INTRODUCTION

Tuberculosis is a global health problem that causes 1. 4 million deaths every year. It has been estimated that sputum smear-negative diagnosis but culture-positive pulmonary TB diagnosis contribute to 12.6% of pulmonary TB transmission. TB diagnosis by smear microscopy smear has a minimum detection limit (LOD) of 5,000 to 10,000 bacilli per milliliter (CFU/ml) of sputum result in missed cases and false positives. However, GeneXpert technology, with a LOD of 131-250 CFU/ml in sputum samples and its implementation is believe to facilitate early detection TB and drug-resistant TB case. Since 2013, Ghana health Service (GHS) introduce GeneXpert MTB/RIF diagnostic in all regional hospitals in Ghana, however no assessment of performance between microscopy and GeneXpert TB diagnosis cross the health facilities has been reported. The study compared the results of routine diagnoses of TB by microscopy and Xpert MTB from 2016 to 2020 at the Cape Coast Teaching Hospital (CCTH).

METHODS

The study compared routine microscopic and GeneXpert TB diagnosis results at the Cape Coast Teaching Hospital (CCTH) from 2016 to 2020 retrospectively. Briefly, sputum specimens were collected into 20 mL sterile screw-capped containers for each case of suspected TB infection and processed within 24 h. The samples were decontaminated using the NALC-NaOH method with the final NaOH concentration of 1%. The supernatants were discarded after the centrifuge and the remaining pellets dissolved in 1-1.5 ml of phosphate buffer saline (PBS) and used for diagnosis. A fixed smears were Ziehl-Neelsen acid-fast stain and observed under microscope and the remainings were used for GeneXpert MTB/RIF diagnosis. The data were analyze using GraphPad Prism.

RESULTS

50.11% (48.48-51.38%) were females with an odd ratio (95% CI) of 1.004 (0.944-1.069) more likely to report to the TB clinic for suspected TB diagnosis. The smear-positive cases for the first sputum were 6.6% (5.98-7.25%), and the second sputum was 6.07% (5.45-6.73%). The Xpert MTB-RIF diagnosis detected 2.93% (10/341) (1.42-5.33%) in the first and 5.44% (16/294) (3.14-8.69%) in the second smear-negative TB samples. The prevalence of Xpert MTB-RIF across smear positive showed that males had 56.87% (178/313) and 56.15% (137/244) and females had 43.13% (135/313) and 43.85% (107/244) for the first and second sputum. Also, false negative smears were 0.18% (10/5607) for smear 1 and 0.31% (16/5126) for smear 2.

CONCLUSION

In conclusion, the study highlights the higher sensitivity of the GeneXpert assay compared to traditional smear microscopy for detecting MTB. The GeneXpert assay identified 10 and 16 positive MTB from smear 1 and smear 2 samples which were microscopic negative.

摘要

介绍

结核病是一个全球性的健康问题,每年导致 140 万人死亡。据估计,痰涂片阴性但培养阳性的肺结核诊断占肺结核传播的 12.6%。通过显微镜检查痰涂片进行肺结核诊断的最低检测限(LOD)为每毫升痰 5000 至 10000 个细菌,因此会导致漏诊和假阳性。然而,GeneXpert 技术在痰样本中的 LOD 为 131-250 CFU/ml,其实施被认为有助于早期发现结核病和耐药结核病病例。自 2013 年以来,加纳卫生服务局(GHS)在加纳所有地区医院引入了 GeneXpert MTB/RIF 诊断,但尚未报告在卫生机构之间进行显微镜和 GeneXpert TB 诊断性能的评估。本研究比较了 2016 年至 2020 年在开普敦教学医院(CCTH)进行的常规显微镜和 Xpert MTB 诊断结果。

方法

本研究回顾性比较了 2016 年至 2020 年开普敦教学医院(CCTH)常规显微镜和 GeneXpert TB 诊断结果。简要地说,对每个疑似结核病感染的病例采集 20 毫升无菌螺旋盖容器中的痰液标本,并在 24 小时内处理。使用 NALC-NaOH 法对标本进行去污处理,最终的 NaOH 浓度为 1%。离心后丢弃上清液,将剩余的沉淀物溶解在 1-1.5ml 磷酸盐缓冲液(PBS)中用于诊断。固定涂片采用齐尔-尼尔森抗酸染色并在显微镜下观察,其余用于 GeneXpert MTB/RIF 诊断。使用 GraphPad Prism 分析数据。

结果

50.11%(48.48-51.38%)为女性,优势比(95%CI)为 1.004(0.944-1.069),更有可能因疑似结核病诊断而到结核病诊所就诊。第一次痰液的涂片阳性病例为 6.6%(5.98-7.25%),第二次痰液为 6.07%(5.45-6.73%)。Xpert MTB-RIF 诊断在第一次和第二次痰涂片阴性的 TB 样本中分别检测到 2.93%(10/341)(1.42-5.33%)和 5.44%(16/294)(3.14-8.69%)。在涂片阳性的 Xpert MTB-RIF 中,男性的第一和第二痰液的阳性率分别为 56.87%(178/313)和 56.15%(137/244),女性的阳性率分别为 43.13%(135/313)和 43.85%(107/244)。此外,第一和第二痰液的假阴性涂片率分别为 0.18%(10/5607)和 0.31%(16/5126)。

结论

总之,该研究强调了 GeneXpert 检测法比传统的显微镜痰涂片检测法在检测 MTB 方面具有更高的敏感性。GeneXpert 检测法从涂片 1 和涂片 2 样本中分别鉴定出了 10 个和 16 个 MTB,这些样本的显微镜检查均为阴性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10a0/11218342/8ffa19538002/12879_2024_9566_Fig1_HTML.jpg

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