The stem-loop RT qPCR assay has been proven to offer high sensitivity and specificity for the quantification of mature miRNAs.
The no-template controls gave non-detectable signals in all samples, confirming the high specificity of the miRNA quantification assay.
>> To demonstrate the specificity of the absolute quantification, we performed a ten-fold dilution for each samples and we confirmed that there was a linearity for each miRNAs between the two samples (fold-dilution average between samples: mir-16-5p: 8,81 ± 0,675, mir-486-5p: 7,67 ± 2,41, mir-660-5p: 9,96 ± 1,76).
A specific standard curve was performed in parallel for each gene to assess the specificity of the products, for quantification of the respective transcripts in duplicate.
The method developed in this study showed sensitivity and specificity in the quantification of pork meat in commercial processed meat products.
Since ccn1 and ccn2 are members of the same gene family, it is critically important to examine the specificity of each quantification system in order to avoid possible cross-recognition [ 4].
The high degree of sequence homology between the genes has led to difficulties in previous studies with the specificity of transcript quantification (23– 23).
Table 2 shows the numerical relationships among sensitivity and specificity and PAR% of the quantification for various thresholds c in Figure 2.
In this study a new method was developed and validated for the absolute quantification of endogenous angiotensin II levels in human plasma, since recent studies demonstrated the insufficient specificity of the monoclonal antibodies using for the quantification of angiotensin II by commercial ELISA based assay [ 6, 7].
Nevertheless, the specificity of the newly constructed ELISA made the quantification of accurate levels of Aβ43 possible (Supporting Information Fig S1).
