Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays

将自动化引入克氏锥虫感染的分子诊断:样品处理、DNA 提取方法和实时 PCR 检测的比较研究

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作者:Alba Abras, Cristina Ballart, Teresa Llovet, Carme Roig, Cristina Gutiérrez, Silvia Tebar, Pere Berenguer, María-Jesús Pinazo, Elizabeth Posada, Joaquim Gascón, Alejandro G Schijman, Montserrat Gállego, Carmen Muñoz

Background

Polymerase chain reaction (PCR) has become a useful tool for the diagnosis of Trypanosoma cruzi infection. The development of automated DNA extraction methodologies and PCR systems is an important step toward the standardization of protocols in routine diagnosis. To date, there are only two commercially available Real-Time PCR assays for the routine laboratory detection of T. cruzi DNA in clinical samples: TCRUZIDNA.CE (Diagnostic Bioprobes Srl) and RealCycler CHAG (Progenie Molecular). Our

Significance

This is the first study to evaluate the RealCycler CHAG assay taking into account the overall process, including three variables (sample treatment, extraction method and Real-Time PCR). Our findings may contribute to the harmonization of protocols between laboratories and to a wider application of Real-Time PCR in molecular diagnostic laboratories associated with health centers.

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