Abstract：
Malaria and babesiosis are global health threats affecting humans, wildlife, and domestic animals, particularly in Africa, the Americas, and Europe. Malaria can lead to severe outcomes, while babesiosis usually resembles a mild illness but can be severe and fatal in individuals with weakened immune systems. Swift, accurate detection of these parasites is crucial for treatment and control. We evaluated a real-time PCR assay for diagnosing five Plasmodium and three Babesia species from blood samples, assessing its sensitivity, specificity, and analytical performance by analyzing 46 malaria-positive and 32 Babesia spp-positive samples diagnosed through microscopy. The limit of detection for Plasmodium species ranged from 30 to 0.0003 copies/µL. For mixed infections, it was 0.3 copies/µL for P. falciparum/P. vivax and 3 copies/µL for P. malariae/P. knowlesi. Babesia species had a detection limit of 0.2 copies/µL. No cross-reactivity was observed among 64 DNA samples from various microorganisms. The assay showed good sensitivity, detecting Plasmodium and Babesia species with 100 % accuracy overall, except for P. falciparum (97.7 %) and B. microti (12.5 %). The low sensitivity of detecting B. microti was attributed to limitations in microscopy for species identification. This technique heavily relies on the proficiency of the examiner, as species within the genus cannot be distinguished under a microscope. Additionally, Babesia can be confused with the early trophozoite stage (ring forms) of Plasmodium parasites. The findings support multiplex qPCR\'s diagnostic superiority over the gold standard, despite higher costs. It offers enhanced sensitivity, specificity, and detects mixed infections, crucial for effective monitoring and diagnosis of malaria and babesiosis in endemic regions with significant public health challenges.
摘要：
疟疾和巴贝西虫病是影响人类的全球健康威胁，野生动物,和家畜，尤其是在非洲，美国,和欧洲。疟疾会导致严重的后果，虽然巴贝斯虫病通常类似于轻度的病毒性疾病，但在免疫系统减弱的个体中可能是严重和致命的。Swift,这些寄生虫的准确检测对于治疗和控制至关重要。我们评估了从血液样本中诊断五种疟原虫和三种巴贝虫的实时PCR检测方法，评估其敏感性，特异性，通过分析通过显微镜诊断的46例疟疾阳性和32例巴贝斯虫阳性样本进行分析。疟原虫的检测限范围为30至0.0003拷贝/μL。对于混合感染，恶性疟原虫/P为0.3拷贝/微升hivax和3个拷贝/µL的malariae/P。Knowlesi.巴贝虫物种的检测限为0.2拷贝/微升。在来自各种微生物的64个DNA样品中未观察到交叉反应性。该测定法显示出异常的灵敏度，检测疟原虫和巴贝虫物种，总体准确率为100%，恶性疟原虫（97.7%）和B.microti（12.5%）除外。检测B.microti的低灵敏度归因于显微镜用于物种鉴定的局限性。这种技术在很大程度上依赖于考官的熟练程度，因为属中的物种在显微镜下无法区分。此外,Babesia可能与疟原虫寄生虫的早期滋养体阶段（环状形式）相混淆。这些发现支持多重qPCR诊断优于金标准，尽管成本较高。它提供了增强的灵敏度，特异性，并检测混合感染，对于在具有重大公共卫生挑战的流行地区有效监测和诊断疟疾和巴贝西虫病至关重要。
