Assessment of droplet digital polymerase chain reaction for measuring BCR-ABL1 in chronic myeloid leukaemia in an international interlaboratory study

Measurement of BCR activator of RhoGEF and GTPase -ABL proto-oncogene 1, non-receptor tyrosine kinase (BCR-ABL1) mRNA levels by reverse transcription quantitative polymerase chain reaction (RTqPCR) has been critical to treatment protocols and clinical trials in chronic myeloid leukaemia; however, interlaboratory variation remains a significant issue. Reverse transcriptase droplet digital PCR (RTddPCR) has shown potential to improve testing but a large-scale interlaboratory study is required to definitively establish this. In the present study, 10 BCR-ABL1-positive samples with levels ranging from molecular response (MR)1·0–MR5·0 were tested by 23 laboratories using RTddPCR with the QXDX BCR-ABL %IS kit. A subset of participants tested the samples using RTqPCR. All 23 participants using RTddPCR detected BCR-ABL1 in all samples to MR4·0. Detection rates for deep-response samples were 95·7% at MR4·5, 78·3% at MR4·7 and 87·0% at MR5·0. Interlaboratory coefficient of variation was indirectly proportional to BCR-ABL1 level ranging from 29·3% to 69·0%. Linearity ranged from 0·9330 to 1·000 (average 0·9936). When results were compared for the 11 participants who performed both RTddPCR and RTqPCR, RTddPCR showed a similar limit of detection to RTqPCR with reduced interlaboratory variation and better assay linearity. The ability to detect deep responses with RTddPCR, matched with an improved linearity and reduced interlaboratory variation will allow improved patient management, and is of particular importance for future clinical trials focussed on achieving and maintaining treatment-free remission

Assessment of genetic diversity in different accessions of Jatropha curcas

The aim of this work was to estimate the genetic diversity among 273 Jatropha curcas L. accessions and to develop a new SNP-based multi-allelic marker assay (Intra-. Locus SNP Haplotype, or LSH) to enhance the genetic discriminatory power of the SNP marker system. The accessions were collected in 15 countries of 3 continents (Africa, Asia and America) and analyzed with SSR, EST-SSR and SNP markers. Cluster analysis revealed the presence of two main genetic groups with an extreme high genetic uniformity and homozygosity of accessions grown in all countries of Africa, Asia, South America, and in some states of Mexico. Differently, an increased genetic variability and heterozygosity was observed in other states of Mexico and Guatemala. The newly developed LSH assay successfully identifies multiple alleles at the same locus and increased the average polymorphic information content of the SNP system. These results confirm the general idea recognizing the Central America region as the center of origin of the species and the genetic homogeneity of the world-wide cultivated accessions, and propose a new powerful SNP-based multi-allelic marker assay for enhancing genetic diversity analysis and selection methods in breeding programs. © 2015 Elsevier B.V