Storage duration and polymerase chain reaction detection of Plasmodium falciparum from blood spots on filter paper
Storage duration and polymerase chain reaction detection of Plasmodium falciparum from blood spots on filter paper
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DOI：
10.4269/ajtmh.2003.69.42
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发表时间：
2003-07-01
影响因子：
3.3
通讯作者：
Patarapotikul, J
中科院分区：
文献类型：
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作者：
Chaorattanakawee, S;Natalang, O;Patarapotikul, J
为评价样本过滤器长期储存对聚合酶链反应（PCR）检测疟疾灵敏度的影响，对252例经显微镜证实的恶性疟原虫疟疾患者的血斑进行了分析，并按储存时间分层。这些斑点是在1996年至2000年期间收集在滤纸上并在室温下储存的。使用基于Chelex的方法提取DNA。出乎意料的是，在第一次纯化后，来自最近储存的样品的PCR的灵敏度较低，并且显示出随着储存时间的推移而逐渐增加（P = 0.003，通过线性趋势的卡方检验）。这表明，PCR抑制剂更容易从最近的血点（< 4岁）比从4岁的血点溶解，从而导致PCR灵敏度的时间依赖性增加。然而，如果再次纯化DNA（当第一次PCR结果为阴性时），累积灵敏度不受储存时间的影响。这表明，只要纯化充分，储存时间不是关键问题。
To evaluate the effect of long-term storage of sample filters on the sensitivity of polymerase chain reaction (PCR) detection of malaria, 252 blood spots from patients with microscopically confirmed Plasmodium falciparum malaria were analyzed and stratified by storage duration. The spots were collected between 1996 and 2000 on filter paper and stored at room temperature. A Chelex-based method was used to extract the DNA. Unexpectedly, after the first purification, the sensitivity of the PCR from recently stored samples was low and showed progressively increased with time storage (P = 0.003, by chi-square test for linear trend). This suggested that PCR inhibitors were easier to dissolve from the more recent blood spots (< 4 years old) than from blood spots 4 years old, thus leading to a time-dependent increase in PCR sensitivity. However, if DNA was purified again (when the first PCR result was negative), the cumulative sensitivity was not influenced by storage duration. This indicated that length of storage is not a critical issue providing purification is sufficient.