Quinn A. Conklin, Dana L. Smith, Guorui Dai, Brandon G. King, Clifford D. Saron, Jue Lin
{"title":"端粒长度测量的远程血液采集：评估样品特性和处理对DNA质量和分析结果的影响","authors":"Quinn A. Conklin, Dana L. Smith, Guorui Dai, Brandon G. King, Clifford D. Saron, Jue Lin","doi":"10.1002/ajhb.70128","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>Telomere length (TL) is a valuable marker of aging and stress that reflects both genetic and environmental influences. Quantitative PCR (qPCR) TL measurement is a powerful and cost-effective assay, especially in population studies with limited quantities of source material. Nevertheless, collecting and transporting high-quality blood samples can be logistically challenging, and research suggests that several preanalytical and analytical factors can influence the reliability and precision of the qPCR assay. Here we describe a procedure for collecting blood remotely in a large-scale study. We then assess the influence of various features of the samples, as well as their collection, transportation, and storage on DNA quality and TL assay outcomes.</p>\n </section>\n \n <section>\n \n <h3> Method</h3>\n \n <p>Participants used at-home collection kits to collect a few drops of whole blood in BD Microtainers during a baseline (<i>n</i> = 265) and 1-year follow-up (<i>n</i> = 178) assessment. DNA was extracted using a magnetic-bead method, and DNA yield, purity, and integrity were assessed. TL was measured using qPCR. To assess inter-assay variation, the coefficient of variation (CV) was calculated across repeated TL measurements (three runs) for each sample. When there was adequate material for duplicate extractions of DNA from the same blood samples, we calculated the intra-class correlation (ICC) of the resultant TL values to assess assay precision.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>Our analyses revealed that as little as 50 μL of blood yielded sufficient DNA for highly precise TL measurement (ICC = 0.962, <i>n</i> = 365). Transportation time and an additional year of storage time at −80°C did not meaningfully affect DNA quality or assay outcomes. However, blood clotting was associated with longer telomere estimates, whereas greater temperature exposure was related to shorter telomere estimates.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>We established that whole blood collected remotely in BD Microtainers can provide a valid sample source for qPCR TL measurement. We also outline important logistical considerations related to sample collection and handling and provide recommendations for researchers who want to use this method.</p>\n </section>\n </div>","PeriodicalId":50809,"journal":{"name":"American Journal of Human Biology","volume":"37 9","pages":""},"PeriodicalIF":1.7000,"publicationDate":"2025-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ajhb.70128","citationCount":"0","resultStr":"{\"title\":\"Remote Blood Collection for Telomere Length Measurement: Assessing the Impact of Sample Characteristics and Handling on DNA Quality and Assay Outcomes\",\"authors\":\"Quinn A. Conklin, Dana L. Smith, Guorui Dai, Brandon G. King, Clifford D. Saron, Jue Lin\",\"doi\":\"10.1002/ajhb.70128\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Background</h3>\\n \\n <p>Telomere length (TL) is a valuable marker of aging and stress that reflects both genetic and environmental influences. Quantitative PCR (qPCR) TL measurement is a powerful and cost-effective assay, especially in population studies with limited quantities of source material. Nevertheless, collecting and transporting high-quality blood samples can be logistically challenging, and research suggests that several preanalytical and analytical factors can influence the reliability and precision of the qPCR assay. Here we describe a procedure for collecting blood remotely in a large-scale study. We then assess the influence of various features of the samples, as well as their collection, transportation, and storage on DNA quality and TL assay outcomes.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Method</h3>\\n \\n <p>Participants used at-home collection kits to collect a few drops of whole blood in BD Microtainers during a baseline (<i>n</i> = 265) and 1-year follow-up (<i>n</i> = 178) assessment. DNA was extracted using a magnetic-bead method, and DNA yield, purity, and integrity were assessed. TL was measured using qPCR. To assess inter-assay variation, the coefficient of variation (CV) was calculated across repeated TL measurements (three runs) for each sample. 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Remote Blood Collection for Telomere Length Measurement: Assessing the Impact of Sample Characteristics and Handling on DNA Quality and Assay Outcomes
Background
Telomere length (TL) is a valuable marker of aging and stress that reflects both genetic and environmental influences. Quantitative PCR (qPCR) TL measurement is a powerful and cost-effective assay, especially in population studies with limited quantities of source material. Nevertheless, collecting and transporting high-quality blood samples can be logistically challenging, and research suggests that several preanalytical and analytical factors can influence the reliability and precision of the qPCR assay. Here we describe a procedure for collecting blood remotely in a large-scale study. We then assess the influence of various features of the samples, as well as their collection, transportation, and storage on DNA quality and TL assay outcomes.
Method
Participants used at-home collection kits to collect a few drops of whole blood in BD Microtainers during a baseline (n = 265) and 1-year follow-up (n = 178) assessment. DNA was extracted using a magnetic-bead method, and DNA yield, purity, and integrity were assessed. TL was measured using qPCR. To assess inter-assay variation, the coefficient of variation (CV) was calculated across repeated TL measurements (three runs) for each sample. When there was adequate material for duplicate extractions of DNA from the same blood samples, we calculated the intra-class correlation (ICC) of the resultant TL values to assess assay precision.
Results
Our analyses revealed that as little as 50 μL of blood yielded sufficient DNA for highly precise TL measurement (ICC = 0.962, n = 365). Transportation time and an additional year of storage time at −80°C did not meaningfully affect DNA quality or assay outcomes. However, blood clotting was associated with longer telomere estimates, whereas greater temperature exposure was related to shorter telomere estimates.
Conclusions
We established that whole blood collected remotely in BD Microtainers can provide a valid sample source for qPCR TL measurement. We also outline important logistical considerations related to sample collection and handling and provide recommendations for researchers who want to use this method.
期刊介绍：
The American Journal of Human Biology is the Official Journal of the Human Biology Association.
The American Journal of Human Biology is a bimonthly, peer-reviewed, internationally circulated journal that publishes reports of original research, theoretical articles and timely reviews, and brief communications in the interdisciplinary field of human biology. As the official journal of the Human Biology Association, the Journal also publishes abstracts of research presented at its annual scientific meeting and book reviews relevant to the field.
The Journal seeks scholarly manuscripts that address all aspects of human biology, health, and disease, particularly those that stress comparative, developmental, ecological, or evolutionary perspectives. The transdisciplinary areas covered in the Journal include, but are not limited to, epidemiology, genetic variation, population biology and demography, physiology, anatomy, nutrition, growth and aging, physical performance, physical activity and fitness, ecology, and evolution, along with their interactions. The Journal publishes basic, applied, and methodologically oriented research from all areas, including measurement, analytical techniques and strategies, and computer applications in human biology.
Like many other biologically oriented disciplines, the field of human biology has undergone considerable growth and diversification in recent years, and the expansion of the aims and scope of the Journal is a reflection of this growth and membership diversification.
The Journal is committed to prompt review, and priority publication is given to manuscripts with novel or timely findings, and to manuscripts of unusual interest.
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