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Aim Next Generation Sequencing (NGS) is rapidly becoming the method of choice for HLA typing for Hematopoietic Stem Cell Transplantation. Jeevan Stem Cell Foundation, Chennai, India validated the full gene HLA typing using Immucor’s MIA FORA FLEX kits from conceptualization to implementation in ninety days. This implementation, in line with the Indian Government’s policy of “Make in India”, enabled Jeevan to establish a cost effective high resolution HLA typing to international standards in India to support the transplant physicians and registries in the region and prevent Indian DNA being shipped to other countries. Methods Reference DNA used for validation were previously typed by SBT included samples from UK NEQAS, FRED HUTCHINSON and Jeevan’s registry samples. DNA was extracted using the Qiacube. HLA typing for A, B, C, DRB1 and DQB1 were performed. All workflows were performed according to the supplier’s instructions. NGS sequencing was done using the mid output flow cell kit from Illumina and sequenced on the Illumina MiniSeq platform. Immucor’s proprietary software was used to analyse the data. Results Results were evaluated per allele on the 2nd field consensus compared to the reference, the number of allelic ambiguities, genotype ambiguities and allelic drop outs were measured. The second field resolution was used because higher resolution typing was not available for all samples. MIA FORA NGS software provided high resolution typing for all samples. There results from NGS typing compared to SBT typing are shown in Table 1. Conclusions The implementation of the MIA FORA FLEX HLA typing kit at Jeevan is the first outside USA. With the MIA FORA kits, the turnaround time will depend upon the number of samples. In our experience reports for 48 samples can be delivered within 72 h. There were no ambiguous results and no allelic dropout. The proprietary software of Immucor was easy to use and significantly reduced the analysis time. Download high-res image (213KB) Download full-size image |
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