Bakkour Sonia, Chafets Daniel M, Wen Li, Muench Marcus O, Telford Sam R, Erwin James L, Levin Andrew E, Self Deanna, Brès Vanessa, Linnen Jeffrey M, Lee Tzong-Hae, Busch Michael P
Blood Systems Research Institute, San Francisco, California.
Department of Laboratory Medicine, University of California, San Francisco, California.
Transfusion. 2018 Dec;58(12):2903-2910. doi: 10.1111/trf.14889. Epub 2018 Sep 28.
Babesia microti is a parasite that infects red blood cells (RBCs) in mammals. It is transmitted to humans by tick bites, transfusion, organ transplantation, and congenital acquisition. Although the Babesia natural history and seroprevalence in donors have been well described, gaps in knowledge relevant to transfusion remain.
Mice were infected with dilutions of parasitized blood to address the minimal infectious dose and the kinetics of parasitemia by quantitative polymerase chain reaction (qPCR) and of antibodies by enzyme immunoassay.
In immunocompetent DBA/2 mice infected with 100 parasitized RBCs (pRBCs) and in immunodeficient NSG mice infected with 63 pRBCs, parasitemia was detectable in five of five mice each. Peak parasitemia up to 2 × 10 pRBCs/mL at 2 to 3 weeks or 5 × 10 pRBCs/mL at 6 weeks was observed for DBA/2 and NSG mice, respectively. Protracted fluctuating parasitemia was observed for 8 months in DBA/2 mice, whereas NSG mice exhibited a high-plateau parasitemia. Antibody titers continued to increase until 6 to 18 weeks in DBA/2 mice and remained high through 6 months. This study also investigated the analytical performance of Babesia assays that detect parasite DNA or RNA using a blinded panel. A Babesia assay targeting parasite RNA was approximately 10-fold more sensitive compared to qPCR targeting DNA.
The mice in this study were highly susceptible to Babesia infection using as few as 1 to 2 log pRBCs and maintained chronic parasitemia. If the infectious dose in human transfusion recipients is comparably low, a highly sensitive assay targeting parasite RNA may safeguard the blood supply, particularly before antibody detection.
微小巴贝斯虫是一种感染哺乳动物红细胞的寄生虫。它通过蜱叮咬、输血、器官移植和先天性感染传播给人类。尽管巴贝斯虫在供体中的自然史和血清流行率已有充分描述,但与输血相关的知识仍存在空白。
用稀释的感染寄生虫的血液感染小鼠,通过定量聚合酶链反应(qPCR)检测寄生虫血症的最小感染剂量和动力学,并用酶免疫测定法检测抗体。
在感染100个感染寄生虫的红细胞(pRBCs)的免疫活性DBA/2小鼠和感染63个pRBCs的免疫缺陷NSG小鼠中,五只小鼠中的每只均检测到寄生虫血症。DBA/2和NSG小鼠分别在2至3周时观察到高达2×10个pRBCs/mL的寄生虫血症峰值,或在6周时观察到5×10个pRBCs/mL的峰值。在DBA/2小鼠中观察到长达8个月的持续波动的寄生虫血症,而NSG小鼠表现出高平台期寄生虫血症。DBA/2小鼠的抗体滴度持续升高直至6至18周,并在6个月内保持高位。本研究还使用盲法检测小组研究了检测寄生虫DNA或RNA的巴贝斯虫检测方法的分析性能。与靶向DNA的qPCR相比,靶向寄生虫RNA的巴贝斯虫检测方法的灵敏度高约10倍。
本研究中的小鼠对巴贝斯虫感染高度敏感,使用低至1至2个对数的pRBCs并维持慢性寄生虫血症。如果人类输血受者的感染剂量同样较低,一种针对寄生虫RNA的高灵敏度检测方法可能会保障血液供应,特别是在抗体检测之前。