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Abstract Targeted panel-based tumor mutation burden (TMB) assays are widely employed to guide immunotherapy for patients with solid tumors. However, the accuracy and consistency of this method can be compromised due to the variability in technical details across different laboratories, particularly in terms of panel size, somatic mutation detection and TMB calculation rules. Currently, systematic evaluations of the impact of these technical factors on existing assays and best practice recommendations remain lacking. We assessed the performance of 50 participating panel-based TMB assays involving 38 unique methods using cell line samples. In silico experiments utilizing TCGA MC3 datasets were performed to further dissect the impact of technical factors. Here we show that the panel sizes beyond 1.04 Mb and 389 genes are necessary for the basic discrete accuracy, as determined by over 40,000 synthetic panels. The somatic mutation detection should maintain a reciprocal gap of recall and precision less than 0.179 for reliable psTMB calculation results. The inclusion of synonymous, nonsense and hotspot mutations could enhance the accuracy of panel-based TMB assay. A 5% variant allele frequency cut-off is suitable for TMB assays using tumor samples with at least 20% tumor purity. In conclusion, this multicenter study elucidates the major technical factors as sources of variability in panel-based TMB assays and proposed comprehensive recommendations for the enhancement of accuracy and consistency. These findings will assist clinical laboratories in optimizing the methodological details through bioinformatic experiments to enhance the reliability of panel-based methods.

Abstract Transverse cracking is a serious problem for semi-rigid base asphalt pavement. The shrinkage of the base course and the surface course, as well as reflective cracks, are key factors for transverse cracking in asphalt pavement. Crack spacing can directly reflect the degree of transverse cracking in pavements. Therefore, this study aims to calculate the transverse crack spacing and discuss its affecting factors. To this end, a calculation model of transverse crack spacing for the semi-rigid base asphalt pavement was first established. Then, the transverse crack spacings of different composite structures were calculated, and the influences of the shrinkage coefficient, the structural layer thickness, and the pavement tensile strength on transverse crack spacing were expounded. Finally, the transverse crack spacing of the pavement after the appearance of the reflective was calculated. The results show that the lower lime and fly ash content and skeleton gap gradation can be adopted during the design of the base course. Meanwhile, the lower lime and fly ash content in the macadam base, the skeleton gap gradation and asphalt concrete with a larger particle size in the surface layer can be used during the design of surface layer. In addition, the transverse crack spacing of the semi-rigid base asphalt pavement could be increased by reducing the shrinkage coefficient, increasing the thicknesses of the surface course and the base course, and improving the tensile strength of the pavement. After the appearance of reflective cracks, the transverse crack spacing of the surface layer ranged between 32.8 m and 66.5 m. 15fp-AC25, 15fp-AC20, 15df-AC25, and 17fp-AC25 were found to be the best semi-rigid base asphalt pavement structures to reduce transverse cracking. Finally, transverse cracking in pavement composite structures under different bonding conditions needs to be analyzed in the follow-up work.