Ranjbaran Mohsen, Kaur Simerdeep, Wang Jiangshan, Raut Bibek, Verma Mohit S
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana, United States of America.
Birck Nanotechnology Center, Purdue University, West Lafayette, Indiana, United States of America.
PLoS One. 2024 Dec 31;19(12):e0315444. doi: 10.1371/journal.pone.0315444. eCollection 2024.
Nucleic-acid biosensors have emerged as useful tools for on-farm detection of foodborne pathogens on fresh produce. Such tools are specifically designed to be user-friendly so that a producer can operate them with minimal training and in a few simple steps. However, one challenge in the deployment of these biosensors is delivering precise sample volumes to the biosensor's reaction sites. To address this challenge, we developed an innovative drop dispenser using advanced 3D printing technology, combined with a hydrophilic surface chemistry treatment. This dispenser enables the generation of precise sample drops, containing DNA or bacterial samples, in volumes as small as a few micro-liters (∼20 to ∼33 μL). The drop generator was tested over an extended period to assess its durability and usability over time. The results indicated that the drop dispensers have a shelf life of approximately one month. In addition, the device was rigorously validated for nucleic acid testing, specifically by using loop-mediated isothermal amplification (LAMP) for the detection of Escherichia coli O157, a prevalent foodborne pathogen. To simulate real-world conditions, we tested the drop dispensers by integrating them into an on-farm sample collection system, ensuring they deliver samples accurately and consistently for nucleic acid testing in the field. Our results demonstrated similar performance to commercial pipettors in LAMP assays, with a limit of detection of 7.8×106 cells/mL for whole-cell detection. This combination of precision, ease of use, and durability make our drop dispenser a promising tool for enhancing the effectiveness of nucleic acid biosensors in the field.
核酸生物传感器已成为用于在农场检测新鲜农产品上食源性病原体的有用工具。此类工具经过专门设计,便于用户操作,这样生产者只需经过最少的培训,通过几个简单步骤就能操作它们。然而,部署这些生物传感器面临的一个挑战是将精确体积的样品输送到生物传感器的反应位点。为应对这一挑战,我们利用先进的3D打印技术,并结合亲水性表面化学处理,开发了一种创新的滴液分配器。这种分配器能够生成精确的样品液滴,其中含有DNA或细菌样品,体积小至几微升(约20至约33微升)。对该滴液发生器进行了长期测试,以评估其随时间的耐用性和可用性。结果表明,滴液分配器的保质期约为一个月。此外,该设备经过了严格的核酸检测验证,特别是通过使用环介导等温扩增技术(LAMP)检测大肠杆菌O157,这是一种常见的食源性病原体。为模拟实际情况,我们将滴液分配器集成到农场样品采集系统中进行测试,确保它们能准确、一致地输送样品用于现场核酸检测。我们的结果表明,在LAMP检测中,其性能与商用移液器相似,全细胞检测的检测限为7.8×106个细胞/毫升。这种精确性、易用性和耐用性的结合,使我们的滴液分配器成为增强核酸生物传感器在现场有效性的一种有前景的工具。