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Cord blood has become an acceptable source of hematopoietic stem cells for transplantation. HLA plays a major role in hematopoietic stem cell transplantation (HSCT). Typing of cord blood samples for HLA alleles has been performed based on the serological and molecular methods. However, with the advent of nextgeneration sequencing technology, HLA typing becomes more accurate and unambiguous (upto intron level). Contamination of cord blood cells with erythropoietic cells poses a challenge in DNA extraction and downstream application. In the present study, DNA extracted from buffy coat of cord blood samples was typed for HLA-A, -B, -C, DRB1, and DQB1 alleles by Illumina miniseq and the sequences were aligned, phased, and mapped by MIA FORA software algorithms. Most frequent alleles found were HLA Afield resolution) HLA typing can be performed on cord blood samples. In order to preserve the precious sample for future use, minimal amount of cord blood samples (postprocessing) could be used for HLA typing purpose. K E Y W O R D S cord blood units, high-throughput typing, HLA typing, NGS

Background Schizophrenia, a chronic severe psychiatric illness of unknown aetiology, has been shown to be associated with HLA alleles but at varied degree in different population. The present study has focussed on analysing the frequency of HLA class I and class II alleles in persons with schizophrenia from South India. Methods Ninety seven individuals with schizophrenia and 103 age‐ and gender‐matched controls were typed for HLA‐ A, B, C, DRB1 and DQB1 loci by next‐generation sequencing in Illumina MiniSeq using MIA FORA NGS FLEX HLA typing kit. Results The results showed that HLA‐A*01:01:01, B*37:01:01 and C*01:02:01 were positively associated with schizophrenia while HLA‐B*35:03:01 and DRB1*04:03:01 were negatively associated. Gender‐specific associations revealed that DRB1*10:01:01 and DQB1*05:01:01 were positively associated while DQB1*03:02:01 was negatively associated with female subjects with schizophrenia. A*24:02:01~B*37:01:01~C*06:02:01~DRB1*10:01:01~DQB1*05:01:01 is the predominant haplotype in schizophrenia population when compared to healthy controls. Amino acid association in susceptible and protective alleles has shown that the presence of peptide in the peptide‐binding groves of mature HLA‐A protein (K, M, V, R and V at 44th, 67th, 150th, 156th and 158th position), HLA‐B protein (D and S at 77th and 99th position) and HLA‐C protein (M at 99th position) confer susceptibility to the disease, only in the absence of E (Glutamic acid) at 74th position in mature HLA‐DRB1 protein. Interaction of amino acids in protective alleles namely B*35:01:01 and DRB1*04:03:01 has revealed that aspartic acid at 114th (D) position in mature HLA‐B protein and glutamic acid (E) at 74th position of mature HLA‐DRB1 protein have a combined effect in protecting against the disease. Conclusion The study has revealed the HLA association with schizophrenia in south Indian population. The amino acid interaction with the disease needs to be confirmed in a larger population.

No abstract

A total of 356 (217 bone marrow and 139 cord blood donors) Malayalam Speaking individuals (Keralites or Malayalis (Here not to be confused with Malayali tribes of Eastern ghats from South India)) from Kerala, Tamilnadu, Karnataka and Telangana were typed for HLA-A, -B, -C, -DRB and -DQB alleles. Out of 356 samples, 253 samples were typed by Sanger sequencing using SeCore Sequencing IVD® kit user protocol (Invitrogen) and the results detected on an automated ABI 3730xl DNA analyzer instrument. This article is protected by copyright. All rights reserved.

The novel HLA‐B allele, HLA‐B*35:03:23, differs from B*35:03:01 by a single‐nucleotide mismatch in exon 1.

Identification of the novel allele HLA‐B*40:379 carrying polymorphisms in an intron, exon and the 3′ untranslated region.

B:C:DRB1:DQB1 allele and haplotype frequencies were determined among Urdu speaking population from South India by Sanger and Next Generation Sequencing. Seventy bone marrow registry donors and 327 cord blood units from the Jeevan Stem Cell Foundation (part of Be The Cure Registry), Chennai, Tamilnadu were included in the study. No overall deviations from expected Hardy-Weinberg equilibrium proportions were observed at all the five loci studied. The most frequent HLA class I alleles observed in this population were A*11:01:01, B*40:06:01, and C*06:02:01, showing a frequency of 15.87%, 12.72%, and 13.22%, respectively. The most frequent class II alleles observed in this population were DRB1*07:01:01 (15.62%) and DQB1*06:01:01 (21.41%). The top ranked haplotype A*01:01:01~B*57:01:01~C*06:02:01~DRB1*07:01:01~DQB1*03:03:02 (2.77%) reported in the present study, was the highly frequent haplotype found in Telugu speaking population who lived in the same region.