Goldman I F, Qari S H, Skinner J, Oliveira S, Nascimento J M, Póvoa M M, Collins W E, Lal A A
Malaria Branch, NCID-CDC, Public Health Service, Atlanta, GA 30333.
Mem Inst Oswaldo Cruz. 1992 Oct-Dec;87(4):583-7. doi: 10.1590/s0074-02761992000400019.
Passage of malaria-infected blood through a two-layered column composed of acid-washed glass beads and CF 11 cellulose removes white cells from parasitized blood. However, because use of glass beads and CF 11 cellulose requires filtration of infected blood separately through these two resins and the addition of ADP, the procedure is time-consuming and may be inappropriate for use in the field, especially when large numbers of blood samples are to be treated. Our modification of this process yields parasitized cells free of contaminating leukocytes, and because of its operational simplicity, large numbers of blood samples can be processed. Our procedure also compares well with those using expensive commercial Sepacell resins in its ability to separate leukocytes from whole blood. As a test of usefulness in molecular biologic investigations, the parasites obtained from the blood of malaria-infected patients using the modified procedure yield genomic DNA whose single copy gene, the circumsporozoite gene, efficiently amplifies by polymerase chain reaction.
让感染疟疾的血液通过由酸洗玻璃珠和CF 11纤维素组成的两层柱,可以从受感染的血液中去除白细胞。然而,由于使用玻璃珠和CF 11纤维素需要分别通过这两种树脂过滤感染的血液并添加ADP,该过程耗时且可能不适用于现场使用,特别是当要处理大量血液样本时。我们对该过程的改进产生了不含污染白细胞的被寄生细胞,并且由于其操作简单,可以处理大量血液样本。我们的方法在从全血中分离白细胞的能力方面也与使用昂贵的商业Sepacell树脂的方法相当。作为在分子生物学研究中有用性的测试,使用改进方法从疟疾感染患者血液中获得的寄生虫产生基因组DNA,其单拷贝基因,即环子孢子蛋白基因,通过聚合酶链反应有效地扩增。