Abstract
Background
The analysis of NPM1 mutations establishes an appropriate molecular genetic parameter for checking the accomplishment of therapy in AML patients, as well as for monitoring the minimal residual disease. The possibility of absolute quantification by means of digital PCR allows quantification without the standard and can be used individually by allele-specific primers.

Methods
The work presents a digital PCR that was established for the purpose of analysis of NPM1 mutations by testing different allele-specific primers and probes.

Results
The testing of the PCR products by gel electrophoresis showed that only the use of the specific reverse primer resulted in no unspecific products. After various optimization steps, the digital PCR was performed at 57 ° C for 40 cycles using 2 ul sample material. Therefore, the sensitivity was determined by testing serial dilutions of 0.01%. As
Ccompared with the method of real-time PCR was ,a significant correlation (Pearson Corr .: 0.97; p <0.001 and 0.92; p <0.05) of the detected results wares determined.

Conclusion
The digital PCR is allowed by a high sensitivity and usability of the individual that monitor the course of therapy, as well as for minimal residual disease by rare mutation types. Consequently, this method help
s to detect/identify a recurrence disease in the early stages, and to so send the patient promptly for an adequate therapy.

The text above was approved for publishing by the original author.

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