随机引物R143在烟曲霉DNA测定中的适用性。

Z. Erjavec, M. Brinker, H. Z. Apperloo-Renkema, J. Arends, H. D. de Vries-Hospers, M. Ruiters
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引用次数: 2

摘要

测定了随机引物R143对烟曲霉DNA的特异性,以检验其在真菌培养物中确定烟曲霉DNA存在的有效性。将这些培养物的PCR反应产物与其他21种细菌和真菌的DNA样品进行比较,R143证明仅与烟曲霉产生1346 bp的条带。该条带已完全测序,EcoRI酶切位点用于后续PCR产物的确认。南方印迹法也证实了烟曲霉DNA的特异性。用R143对培养物中分离的DNA进行PCR分型比较,24例中有22例的形态分型一致。在两种情况下存在不一致:两次PCR结果都正确显示烟曲霉的存在,最初没有通过培养检测到。R143是烟曲霉特异性随机引物,具有用于临床标本烟曲霉DNA检测的潜力。
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Applicability of random primer R143 for determination of Aspergillus fumigatus DNA.
The specificity of random primer R143 for Aspergillus fumigatus DNA was determined in order to test its usefulness in establishing the presence of A. fumigatus DNA in fungal cultures. When PCR reaction products of these cultures were compared with those of 21 other bacterial and fungal DNA samples, R143 proved to produce a 1346 bp band with only A. fumigatus. This band has been sequenced completely and the EcoRI restriction site was used for subsequent confirmation of PCR products. The specificity for A. fumigatus DNA was also confirmed by Southern blotting. Comparison of morphological typing of Aspergillus species in cultures with PCR using R143 on DNA isolated from these cultures showed concordance in 22 of 24 cases. In two cases there was discordance: both times PCR results showed correctly the presence of A. fumigatus, initially not detected by culture. R143 is an A. fumigatus specific random primer, with potential for use in detection of A. fumigatus DNA in clinical specimens.
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