AmplideX® PCR/CE FMR1

AmplideX PCR/CE FMR1 Kit has created an easy-to-use, accessible, high performance method for laboratories to reliably analyse CGG repeats and detect interrupting AGG sequences in the FMR1 gene.

Reduced Complexity | Ease-of-analysis of the FMR1 gene has been simplified through:

• Implementation of proprietary PCR solution for amplifying GC-rich regions
• Automation of result calling using AmplideX PCR/CE FMR1 Reporter

Optimized Workflow | Valuable operator hands-on time has been significantly reduced through:

• Direct injection of PCR products (no PCR clean up) in to Capillary Electrophoresis platforms
• Decreased need for Southern blot analysis (up to 50 fold)
• End-to-end solution for FMR1 analysis including all necessary reagents and software

Quality Performance | Performing FMR1 Analysis with Greater Sensitivity and Accuracy:

• Detection of all allele expansions, including low abundance full mutation size mosaics with up to at least 1300 CGG repeats
• Up to 875 fold more sensitive than Southern blot1
• Resolution of female homozygous and heterozygous samples and indication of interrupting AGG sequences
• Proven performance as indicated by more than 30 peer reviewed publications

Analytical Characteristics of AmplideX PCR/CE FMR1 Kit Include:

• Proven clinical accuracy compared to Southern Blot (Table 1)
• Detects all alleles including low abundance full mutations (Figure 1)
• Accurately sizes all alleles up to 200 CGG repeats (Figure 2)
• Resolves female zygosity (Figure 3)
• Detects presence of AGG interruptions (Figure 4)

Table 1: Diagnostic Sensitivity of 100%; Diagnostic Specificity of 98.4% and Overall Accuracy of 99%

*These 2 samples presented premutation alleles by both methods and low intensity full mutation alleles detected only by the AmplideX PCR/CE FMR1 Kit

Figure 1: Amplification of Asuragen’s Methylation and Sensitivity Control which has a 5% full mutation in a background of 95% Normal

Figure 2. Female Pre-mutation sample with accurate sizing of Normal (30 CGG) and Pre-mutation allele (56 CGG)

Figure 3: The difference in the “stutter” peak patterns of homozygous and heterozygous female provides a clear resolution of zygosity

Figure 4. Female Full Mutation sample with AGG interruptions as indicated by sudden decrease in peak heights of the “stutter” peak profile