Abstract：
:Automated extraction of DNA for testing of laboratory samples is an attractive alternative to labour-intensive manual methods when higher throughput is required. However, it is important to maintain the maximum detection sensitivity possible to reduce the occurrence of type II errors (false negatives; failure to detect the target when it is present), especially in the biomedical field, where PCR is used for diagnosis. We used blood infected with known concentrations of Trypanosoma copemani to test the impact of analysis techniques on trypanosome detection sensitivity by PCR. We compared combinations of a manual and an automated DNA extraction method and two different PCR primer sets to investigate the impact of each on detection levels. Both extraction techniques and specificity of primer sets had a significant impact on detection sensitivity. Samples extracted using the same DNA extraction technique performed substantially differently for each of the separate primer sets. Type I errors (false positives; detection of the target when it is not present), produced by contaminants, were avoided with both extraction methods. This study highlights the importance of testing laboratory techniques with known samples to optimise accuracy of test results.
journal_name
Exp Parasitoljournal_title
Experimental parasitologyauthors
Dunlop J,Thompson CK,Godfrey SS,Thompson RCdoi
10.1016/j.exppara.2014.08.006subject
Has Abstractpub_date
2014-11-01 00:00:00pages
20-4eissn
0014-4894issn
1090-2449pii
S0014-4894(14)00196-9journal_volume
146pub_type
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journal_title：Experimental parasitology
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journal_title：Experimental parasitology
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