Some key considerations regarding the design and evaluation of high-throughput sequencing-based biallelic chimerism assays

Abstract

With interest I read paper by Li et (2022) [1] on development of a chimerism 12 monitoring assay using next-generation sequencing (NGS) of single nucleotide polymorphisms 13 (SNPs). Such assays have garnered an increasing amount of attention over the last years. 14 Indeed, comparable assays have been proposed and studied in, at least, seven distinct studies 15 (summarized in [2]). Unfortunately, Li et al. do not refer to most of these studies and thus 16 performance, novelty, strengths, limitations or any other important aspect of Li et al.'s assay in 17 relation to a majority of those already proposed remain undiscussed. Even more unfortunate, 18 several claims by Li et al. remain without profound substantiation. Important considerations 19 and parameters remain unreported on, all in an era where arguably a considerable proportion of 20 science suffers a replicability crisis. 21 An informativity rate of 99.99% is distilled out of thin air. Informativity rates of NGS-based 22 chimerism assays have been reported using empirical data [3]. Admittedly very recently, we 23 have developed a framework that allows calculation of the informativity rate of biallelic 24