Analysis of malaria parasite RNA from decade-old Giemsa-stained blood smears and dried mosquitoes
Analysis of malaria parasite RNA from decade-old Giemsa-stained blood smears and dried mosquitoes
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DOI：
10.4269/ajtmh.1997.57.727
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发表时间：
1997-12-01
影响因子：
3.3
通讯作者：
McCutchan, TF
中科院分区：
文献类型：
--
作者：
Li, J;Wirtz, RA;McCutchan, TF
我们分析了从最近制备的和历史上保存的含有疟原虫感染血液涂片的载玻片中分离的RNA。我们发现保存长达20年的载玻片可以产生RNA，这是一种适合聚合酶链反应（PCR）扩增的模板。在环境温度下储存多年的蚊子也可以用作RNA来源。从载玻片衍生的材料中扩增出的RNA依赖于逆转录酶的添加，其产物对寄生虫的发育状态是特异性的。用疟原虫保守的引物扩增核糖体RNA，并用物种特异性探针杂交，为物种测定提供了一种通用的、无偏的方法。来自幻灯片的信使RNA转录物似乎也可以作为模板。该程序可以为吉姆萨染色血涂片的显微检查提供补充信息，包括物种鉴定、变异抗原鉴定和耐药性状况。
We have analyzed RNA isolated from recently prepared and historically preserved slides containing smears of Plasmodium-infected blood. We found that slides preserved as long as 20 years can yield RNA that is a suitable template for polymerase chain reaction (PCR) amplification. Mosquitoes that have been stored for years under ambient temperature can also be used as an RNA source. The RNA amplification from slide-derived material is shown to be dependent upon the addition of reverse transcriptase and the resultant products are specific to the developmental state of the parasite. Amplification of ribosomal RNA with primers conserved for Plasmodium and hybridization with species-specific probes provide a general, unbiased method for species determination. Messenger RNA transcripts from slides also appear to serve as templates. The procedure may add complementary information to that derived from microscopic examination of Giemsa-stained blood smears including species identification, variant antigen identification and drug resistance status.