两种 PCR 方案在鉴别卵形疟原虫种间差异的比较及其对返回德国旅行者临床管理的意义：一项为期 10 年的横断面研究。
A comparison of two PCR protocols for the differentiation of Plasmodium ovale species and implications for clinical management in travellers returning to Germany: a 10-year cross-sectional study.
机构信息
Department of Microbiology and Hospital Hygiene, External Site at the Bernhard Nocht Institute, Tropical Microbiology and Entomology, Bundeswehr Hospital Hamburg, Bernhard Nocht Str. 74, 20359, Hamburg, Germany.
Institute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, Rostock, Germany.
出版信息
Malar J. 2019 Aug 9;18(1):272. doi: 10.1186/s12936-019-2901-0.
BACKGROUND
To assess the occurrence of Plasmodium ovale wallikeri and Plasmodium ovale curtisi species in travellers returning to Germany, two real-time PCR protocols for the detection and differentiation of the two P. ovale species were compared. Results of parasite differentiation were correlated with patient data.
METHODS
Residual nucleic acid extractions from EDTA blood samples of patients with P. ovale spp. malaria, collected between 2010 and 2019 at the National Reference Centre for Tropical Pathogens in Germany, were subjected to further parasite discrimination in a retrospective assessment. All samples had been analysed by microscopy and by P. ovale spp.-specific real-time PCR without discrimination on species level. Two different real-time PCR protocols for species discrimination of P. o. curtisi and P. o. wallikeri were carried out. Results were correlated with patient data on gender, age, travel destination, thrombocyte count, and duration of parasite latency.
RESULTS
Samples from 77 P. ovale spp. malaria patients were assessed, with a male:female ratio of about 2:1 and a median age of 30 years. Parasitaemia was low, ranging from few visible parasites up to 1% infected erythrocytes. Discriminative real-time PCRs revealed 41 cases of P. o. curtisi and 36 cases of P. o. wallikeri infections. Concordance of results by the two PCR approaches was 100%. Assessment of travel destinations confirmed co-existence of P. o. curtisi and P. o. wallikeri over a wide range of countries in sub-Saharan Africa. Latency periods for the two P. ovale species were similar, with median values of 56.0 days for P. o. curtisi and 58.0 days for P. o. wallikeri; likewise, there was no statistically significant difference in thrombocyte count with median values of 138.5/µL for patients with P. o. curtisi and 152.0/µL for P. o. wallikeri-infected patients.
CONCLUSIONS
Two different real-time PCR protocols were found to be suitable for the discrimination of P. o. curtisi and P. o. wallikeri with only minor differences in sensitivity. Due to the overall low parasitaemia and the lack of differences in severity-related aspects like parasite latency periods or thrombocyte counts, this study supports the use of P. ovale spp. PCR without discrimination on species level to confirm the diagnosis and to inform clinical management of malaria in these patients.
背景
为了评估返回德国的旅行者中卵形疟原虫壁克里亚种和卵形疟原虫库蒂斯亚种的发生情况，比较了两种用于检测和区分这两种卵形疟原虫的实时 PCR 方案。寄生虫分化的结果与患者数据相关。
方法
对 2010 年至 2019 年间在德国热带病国家参考中心采集的卵形疟原虫 spp.疟疾患者的 EDTA 血液样本中的残留核酸进行进一步的寄生虫鉴别，进行回顾性评估。所有样本均通过显微镜检查和卵形疟原虫 spp.特异性实时 PCR 进行分析，而不区分种属。进行了两种不同的实时 PCR 方案来区分卵形疟原虫 curtisi 和卵形疟原虫 wallikeri。结果与患者的性别、年龄、旅行目的地、血小板计数和寄生虫潜伏期相关联。
结果
对 77 例卵形疟原虫 spp.疟疾患者的样本进行了评估，男性与女性的比例约为 2:1，中位年龄为 30 岁。寄生虫血症较低，范围从少数可见寄生虫到 1%感染的红细胞。有区别的实时 PCR 显示 41 例卵形疟原虫 curtisi 感染和 36 例卵形疟原虫 wallikeri 感染。两种 PCR 方法的结果一致性为 100%。对旅行目的地的评估证实，卵形疟原虫 curtisi 和卵形疟原虫 wallikeri 广泛存在于撒哈拉以南非洲的多个国家。两种卵形疟原虫的潜伏期相似，卵形疟原虫 curtisi 的中位值为 56.0 天，卵形疟原虫 wallikeri 的中位值为 58.0 天；同样，血小板计数也没有统计学上的显著差异，卵形疟原虫 curtisi 患者的中位值为 138.5/µL，卵形疟原虫 wallikeri 患者的中位值为 152.0/µL。
结论
发现两种不同的实时 PCR 方案适用于卵形疟原虫 curtisi 和卵形疟原虫 wallikeri 的区分，其敏感性仅有微小差异。由于总体寄生虫血症较低，且潜伏期或血小板计数等与严重程度相关的方面没有差异，因此本研究支持使用不区分种属的卵形疟原虫 spp. PCR 来确认诊断并为这些患者的疟疾临床管理提供信息。
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