Tufa Tafese Beyene, Postigo-Hidalgo Ignacio, Fuchs André, Orth Hans Martin, Häussinger Dieter, Luedde Tom, Kaiser Marco, Patel Pranav, Landt Olfert, Drexler Jan Felix, Feldt Torsten
Hirsch Institute of Tropical Medicine, Asella, Ethiopia.
College of Health Sciences, Arsi University, Asella, Ethiopia.
Am J Trop Med Hyg. 2024 Nov 12;112(1):79-84. doi: 10.4269/ajtmh.23-0663. Print 2025 Jan 8.
Febrile illnesses contribute significantly to morbidity and mortality in sub-Saharan Africa, but the lack of diagnostic facilities and the broad spectrum of pathogens can lead to inadequate clinical management. The timely and reliable identification of the causative pathogens in febrile patients is the basis for the administration of optimal treatment. We aimed to evaluate the performance of a multiplex polymerase chain reaction (PCR) among blood culture-negative patients presenting with febrile diseases in Central Ethiopia. From April 2016 to June 2018, we collected blood samples from adults and children ≥1 year of age admitted with febrile diseases to the Asella Referral and Teaching Hospital, which is located at an altitude of 2,400 m. Total nucleic acids were extracted from frozen plasma samples using a MagNA Pure 96 instrument (Roche, Mannheim, Germany). The multiplex PCR assays were used in combination with LightCycler multiplex DNA master mix (Roche) on a LightCycler 480 instrument (Roche). We used the pathogen-specific assays targeted to Plasmodium spp., Borrelia spp., Rickettsia spp., Leptospira spp., Salmonella spp., and arboviruses. We tested plasma samples of 511 patients and found positive results for Plasmodium spp. (13, 2.5%), Borrelia spp. (12, 2.3%), and Rickettsia species (7, 1.3%); in total, pathogens were detected in 32 of the samples (6.3%). No pathogen was detected by multiplex PCR in 94% of blood culture-negative samples. Even if the pathogens identified by PCR were not necessarily causes of fever, molecular testing using a multiplex PCR can contribute to pathogen diagnosis in a proportion of febrile patients in the highland part of Ethiopia and help to improve the clinical management.
发热性疾病在撒哈拉以南非洲地区的发病率和死亡率中占很大比例，但由于缺乏诊断设施以及病原体种类繁多，可能导致临床治疗不足。及时、可靠地鉴定发热患者的致病病原体是进行最佳治疗的基础。我们旨在评估多重聚合酶链反应（PCR）在埃塞俄比亚中部患有发热性疾病且血培养阴性的患者中的表现。2016年4月至2018年6月，我们从海拔2400米的阿塞拉转诊和教学医院收治的患有发热性疾病的1岁及以上成人和儿童中采集血样。使用MagNA Pure 96仪器（德国曼海姆罗氏公司）从冷冻血浆样本中提取总核酸。多重PCR检测与LightCycler多重DNA主混合液（罗氏公司）联合用于LightCycler 480仪器（罗氏公司）上。我们使用针对疟原虫属、疏螺旋体属、立克次体属、钩端螺旋体属、沙门氏菌属和虫媒病毒的病原体特异性检测方法。我们检测了511名患者的血浆样本，发现疟原虫属（13例，2.5%）、疏螺旋体属（12例，2.3%）和立克次体属（7例，1.3%）呈阳性结果；总共在32个样本（6.3%）中检测到病原体。在94%的血培养阴性样本中，多重PCR未检测到病原体。即使PCR鉴定出的病原体不一定是发热的原因，但使用多重PCR进行分子检测可以帮助埃塞俄比亚高地部分发热患者进行病原体诊断，并有助于改善临床治疗。
