The best practice for preparation of samples from FTA®cards for diagnosis of blood borne infections using African trypanosomes as a model system.
The best practice for preparation of samples from FTA®cards for diagnosis of blood borne infections using African trypanosomes as a model system.
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DOI：
10.1186/1756-3305-4-68
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发表时间：
2011-05-07
影响因子：
3.2
通讯作者：
Picozzi K
中科院分区：
文献类型：
--
作者：
Ahmed HA;MacLeod ET;Hide G;Welburn SC;Picozzi K
血源性传染病的诊断主要依赖于血液样本中病原体的检测。分子技术为此提供了敏感和特定的工具，尽管在现场环境中使用这些方法存在相当大的困难。在大规模流行病学研究中，FTA®卡片越来越受欢迎，用于快速收集和存档大量样本。然而，在这些卡片的下游处理中存在一些困难，这对于准确诊断感染至关重要。在这里，我们描述了使用PCR进行锥虫病分子诊断的FTA®卡样品处理的最佳实践方法的建议。对五种技术进行了比较。直接使用全血的检测灵敏度（35.6%）低于随后使用Chelex®100从卡片中洗脱的全血（56.4%）。在FTA卡片上应用前进行血液溶解时，获得了更好的表观灵敏度（73.3%），尽管这并不显著。随后使用Chelex®100进行洗脱（73.3%）并没有改善这一结果，与现场直接从血液中提取DNA（68.3%）没有显著差异。基于这些结果，每种技术所需的努力程度以及在现场条件下提取DNA的难度，我们建议将血液完整地转移到FTA卡上，然后在Chelex®100中洗脱，这是最佳方法。
Diagnosis of blood borne infectious diseases relies primarily on the detection of the causative agent in the blood sample. Molecular techniques offer sensitive and specific tools for this although considerable difficulties exist when using these approaches in the field environment. In large scale epidemiological studies, FTA®cards are becoming increasingly popular for the rapid collection and archiving of a large number of samples. However, there are some difficulties in the downstream processing of these cards which is essential for the accurate diagnosis of infection. Here we describe recommendations for the best practice approach for sample processing from FTA®cards for the molecular diagnosis of trypanosomiasis using PCR. A comparison of five techniques was made. Detection from directly applied whole blood was less sensitive (35.6%) than whole blood which was subsequently eluted from the cards using Chelex®100 (56.4%). Better apparent sensitivity was achieved when blood was lysed prior to application on the FTA cards (73.3%) although this was not significant. This did not improve with subsequent elution using Chelex®100 (73.3%) and was not significantly different from direct DNA extraction from blood in the field (68.3%). Based on these results, the degree of effort required for each of these techniques and the difficulty of DNA extraction under field conditions, we recommend that blood is transferred onto FTA cards whole followed by elution in Chelex®100 as the best approach.
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影响因子：
3.2
作者：
Cox AP;Tosas O;Tilley A;Picozzi K;Coleman P;Hide G;Welburn SC
通讯作者：
Welburn SC
影响因子：
2.7
作者：
deAlmeida, PJLP;Ndao, M;Geerts, S
通讯作者：
Geerts, S
影响因子：
2.6
作者：
Mahama, CI;Desquesnes, M;Geerts, S
通讯作者：
Geerts, S
影响因子：
2.4
作者：
MOSER, DR;COOK, GA;DONELSON, JE
通讯作者：
DONELSON, JE
影响因子：
2.6
作者：
Delespaux, V;Ayral, F;Geerts, S
通讯作者：
Geerts, S