Identification of 8-Digit HLA-A, -B, -C, and -DRB1Allele and Haplotype Frequencies in Koreans Using the One Lambda AllType Next-Generation Sequencing Kit

Background@#Recent studies have successfully implemented next-generation sequencing (NGS) in HLA typing. We performed HLA NGS in a Korean population to estimate HLA-A, -B, -C, and -DRB1 allele and haplotype frequencies up to an 8-digit resolution, which might be useful for an extended application of HLA results. @*Methods@#A total of 128 samples collected from healthy unrelated Korean adults, previously subjected to Sanger sequencing for 6-digit HLA analysis, were used. NGS was performed for HLA-A, -B, -C, and -DRB1 using the AllType NGS kit (One Lambda, West Hills, CA, USA), Ion Torrent S5 platform (Thermo Fisher Scientific, Waltham, MA, USA), and Type Steam Visual NGS analysis software (One Lambda). @*Results@#Eight HLA alleles showed frequencies of ≥ 10% in the Korean population, namely, A*24:02:01:01 (19.5%), A*33:03:01 (15.6%), A*02:01:01:01 (14.5%), A*11:01:01:01 (13.3%), B*15:01:01:01 (10.2%), C*01:02:01 (19.9%), C*03:04:01:02 (11.3%), and DRB1*09:01:02 (10.2%). Nine previous 6-digit HLA alleles were further identified as two or more 8-digit HLA alleles. Of these, eight alleles (A*24:02:01, B*35:01:01, B*40:01:02, B*55:02:01, B*58:01:01, C*03:02:02, C*07:02:01, and DRB1*07:01:01) were identified as two 8-digit HLA alleles, and one allele (B*51:01:01) was identified as three 8-digit HLA alleles. The most frequent four-loci haplotype was HLA-A*33:03:01-B*44:03:01:01-C*14:03-DRB1*13:02:01. @*Conclusions@#We identified 8-digit HLA-A, -B, -C, and -DRB1 allele and haplotype frequencies in a healthy Korean population using NGS. These new data can be used as a representative Korean data for further disease-related HLA type analysis.

Genetic Polymorphism of CYP2D6 and Clomiphene Concentrations in Infertile Patients with Ovulatory Dysfunction Treated with Clomiphene Citrate

CYP2D6 is primarily responsible for the metabolism of clomiphene citrate (CC). The purpose of the present study was to investigate the relationship between CYP2D6 genotypes, concentrations of CC and its major metabolites and drug response in infertility patients. We studied 42 patients with ovulatory dysfunction treated with only CC. Patients received a dose of 100 mg/day CC on days 3-7 of the menstrual cycle. CYP2D6 genotyping and measurement of CC and the major metabolite concentrations were performed. Patients were categorized into CC responders or non-responders according to one cycle response for the ovulation. Thirty-two patients were CC responders and 10 patients were non-responders with 1 cycle treatment. The CC concentrations were highly variable within the same group, but non-responders revealed significantly lower (E)-clomiphene concentration and a trend of decreased concentrations of active metabolites compared to the responders. Nine patients with intermediate metabolizer phenotype were all responders. We confirmed that the CC and the metabolite concentrations were different according to the ovulation status. However, our results do not provide evidence for the contribution of CYP2D6 polymorphism to either drug response or CC concentrations.

Performance Evaluation of Anyplex plus MTB/NTM and AdvanSure TB/NTM for the Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria / 대한임상미생물학회지

BACKGROUND: Polymerase chain reaction (PCR) methods from direct specimen are widely used for the rapid and accurate detection of mycobacteria infection. In this study, we evaluated two domestically developed detection kits for Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) using real-time PCR. METHODS: A total of 348 samples from patients with suspected tuberculosis were tested with real-time PCR over seven months. We performed real-time PCR using the recently developed Anyplex plus MTB/NTM Detection kit (Seegene) with the CFX 96TM Realtime PCR System (Bio-Rad Laboratories) and the conventional AdvanSure TB/NTM real-time PCR kit (LG Life Sciences) with the SLAN Real-time PCR detection system (LG Life Sciences) to evaluate their performance for detecting MTB and NTM. RESULTS: The two real-time PCR systems showed 96.8% concordance rate for MTB-positive, NTM-positive, and negative results. Based on culture results, the sensitivity and specificity for the detection of MTB using PCR were 71.0% and 94.9% for Anyplex plus, and 78.1% and 93.9% for AdvanSure, respectively. For the detection of NTM, the sensitivity and specificity were 33.3% and 98.4% for Anyplex plus, and 51.7% and 97.9% for AdvanSure. Both PCR systems showed high MTB positive results in bronchial washing and sputum samples. CONCLUSION: In detecting MTB and NTM, Anyplex plus MTB/NTM (Seegene) and AdvanSure TB/NTM real-time PCR (LG Life Sciences) showed high concordance rate with each other in all samples. Therefore both detection kits can be used as rapid and reliable detection tool for MTB.