在埃塞俄比亚农村地区的一家医院中，利用显微镜和分子生物学进行疟疾的诊断和评估。
Microscopy and molecular biology for the diagnosis and evaluation of malaria in a hospital in a rural area of Ethiopia.
机构信息
University Institute of Tropical Diseases and Public Health of the Canary Islands University of La Laguna, Tenerife, Spain.
出版信息
Malar J. 2012 Jun 13;11:199. doi: 10.1186/1475-2875-11-199.
BACKGROUND
Malaria is a leading public health problem in Ethiopia. Accurate diagnosis of Plasmodium infections is crucial for the reduction of malaria in tropical areas and for epidemiological studies. The role of light microscopy (LM) as gold standard has been questioned and, therefore, new molecular methods have been developed for the detection of Plasmodium species. The aim of the present work was to compare different malaria diagnostic methods in order to detect the most common species of Plasmodium and to broaden the knowledge of malaria prevalence in a hospital in a rural area in Ethiopia.
METHODS
A cross-sectional survey of 471 individuals was carried out in a hospital in the rural area of Gambo (Ethiopia). Blood samples were prepared for microscopic observation and collected in filter paper for Seminested-Multiplex PCR (SnM-PCR) and real time PCR (qPCR) testing. The SnM-PCR was considered as the gold standard technique and compared with the rest. Thus, agreement between SnM-PCR and LM was determined by calculating Kappa Statistics and correlation between LM and qPCR quantification was calculated by pair-wise correlation co-efficient.
RESULTS
Samples analysed by LM and SnM-PCR were positive for Plasmodium sp. 5.5% and 10.5%, respectively. Sensitivity was 52.2% by LM and 70% by qPCR. Correlation co-efficient between microscopy counts and qPCR densities for Plasmodium vivax was R2 = 0.586. Prevalence was estimated at 7% (95% CI: 4.7-9.3). Plasmodium vivax was the dominant species detected and the difference was statistically significant (χ2 = 5.121 p < 0.05). The highest prevalence of the parasite (10.9%) was observed in age groups under 15 years old.
CONCLUSION
Accurate malaria diagnostic methods have a great effect in the reduction of the number of malaria-infected individuals. SnM-PCR detection of malaria parasites may be a very useful complement to microscopic examination in order to obtain the real prevalence of each Plasmodium species. Although SnM-PCR shows that it is a good tool for the determination of Plasmodium species, today light microscopy remains the only viabletool for malaria diagnosis in developing countries. Therefore, re-inforcement in the training of microscopists is essential for making the correct diagnosis of malaria. Plasmodium vivax was the predominant species in Gambo, a meso-endemic area for this species.
背景
疟疾是埃塞俄比亚的一个主要公共卫生问题。准确诊断疟原虫感染对于减少热带地区的疟疾和进行流行病学研究至关重要。作为金标准的显微镜检查（LM）的作用受到了质疑，因此，已经开发出了新的分子方法来检测疟原虫物种。本研究的目的是比较不同的疟疾诊断方法，以检测最常见的疟原虫物种，并扩大对埃塞俄比亚农村地区一家医院疟疾流行情况的了解。
方法
对农村甘博医院的 471 人进行了横断面调查。准备显微镜观察的血样，并收集在滤纸中进行 Seminested-Multiplex PCR（SnM-PCR）和实时 PCR（qPCR）检测。SnM-PCR 被认为是金标准技术，并与其他技术进行比较。因此，通过计算 Kappa 统计量来确定 SnM-PCR 与 LM 的一致性，并通过两两相关系数来计算 LM 与 qPCR 定量之间的相关性。
结果
通过 LM 和 SnM-PCR 分析的样本分别对疟原虫 sp.呈阳性，阳性率分别为 5.5%和 10.5%。LM 的敏感性为 52.2%，qPCR 的敏感性为 70%。间日疟原虫镜检计数与 qPCR 密度之间的相关系数为 R2=0.586。疟疾病例的患病率估计为 7%（95%CI：4.7-9.3）。间日疟原虫是检测到的主要物种，差异具有统计学意义（χ2=5.121，p<0.05）。寄生虫的最高患病率（10.9%）出现在 15 岁以下年龄组。
结论
准确的疟疾诊断方法对减少疟疾病例具有重要影响。SnM-PCR 检测疟原虫可能是一种非常有用的显微镜检查补充方法，可获得每种疟原虫的真实流行率。尽管 SnM-PCR 表明它是确定疟原虫物种的良好工具，但目前在发展中国家，显微镜检查仍然是唯一可行的疟疾诊断工具。因此，加强对显微镜检查人员的培训对于正确诊断疟疾至关重要。间日疟原虫是甘博的主要物种，该地区为该物种的中间流行区。
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