Chiarella Pieranna, Sisto Renata, de Marco Ario
INAIL Research - Department of Occupational & Environmental Medicine, Epidemiology & Hygiene. Via Fontana Candida 1 - 00078 Monte Porzio Catone (RM), Italy.
Laboratory for Environmental & Life Sciences - University of Nova Gorica, Vipavska 13, S-5000 Nova Gorica, Slovenia.
Future Sci OA. 2018 Dec 11;5(2):FSO361. doi: 10.4155/fsoa-2018-0075. eCollection 2019 Feb.
Gene polymorphism biomarkers identify individual susceptibility to environmental and occupational hazards. The conventional approach considers polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis (RFLP), a reliable but expensive and time-consuming two-step procedure. Therefore we evaluated the simpler method confronting two-pair primers (CTPP)-PCR for its robustness and applicability to epidemiologic studies.
MATERIALS & METHODS: We compared CTPP-PCR and PCR-RFLP techniques to detect two polymorphisms in a set of biological samples.
CTPP-PCR produced contradictory results and required the orthogonal technique for confirming the data.
In contrast to PCR-RFLP, CTPP-PCR of polymorphisms resulted in ambiguous genotyping which strongly jeopardized heterozygosis classification. The necessity of long optimization and control procedures nullified the potential advantages of CTPP-PCR in terms of costs and time.
基因多态性生物标志物可识别个体对环境和职业危害的易感性。传统方法是先进行聚合酶链反应（PCR），然后进行限制性片段长度多态性分析（RFLP），这是一个可靠但昂贵且耗时的两步程序。因此，我们评估了更简单的双引物对（CTPP）-PCR方法的稳健性及其在流行病学研究中的适用性。
我们比较了CTPP-PCR和PCR-RFLP技术，以检测一组生物样本中的两种多态性。
CTPP-PCR产生了矛盾的结果，需要采用正交技术来确认数据。
与PCR-RFLP相比，多态性的CTPP-PCR导致基因分型不明确，这严重危及杂合子分类。长时间的优化和控制程序的必要性抵消了CTPP-PCR在成本和时间方面的潜在优势。
