Abstract：
BACKGROUND: Neonatal sepsis, often attributed to Group B Streptococcus (GBS) infection, poses a critical health risk to infants, demanding rapid and accurate diagnostic approaches. Existing diagnostic approaches are dependent on traditional culture methods, a process that requires substantial time and has the potential to delay crucial therapeutic assessments.
METHODS: This study introduces an innovative Loop-Mediated Isothermal Amplification (LAMP) assay for the early on-site detection of GBS infection from neonatal sepsis blood samples. To develop a LAMP assay, the primers are designed for the selective targeting of a highly conserved segment within the cfb gene encoding the CAMP factor in Streptococcus agalactiae ensuring high specificity.
RESULTS: Rigorous optimization of reaction conditions, including temperature and incubation time, enhances the efficiency of the LAMP assay, enabling rapid and reliable GBS detection within a short timeframe. The diagnostic efficacy of the LAMP assay was evaluated using spiked blood samples by eliminating the DNA extraction step. The simplified colorimetric LAMP assay has the capability to detect S. agalactiae in a neonatal blood sample containing 2 CFU/mL during sepsis. Additionally, the LAMP assay effectively detected S. agalactiae in both the standard and spiked blood samples, with no detectable interference with blood.
CONCLUSIONS: This optimised LAMP assay emerges as a promising tool for early GBS detection, offering a rapid and accurate on-site solution that has the potential to inform timely interventions and improve outcomes in neonatal sepsis cases.
METHODS: This study introduces an innovative Loop-Mediated Isothermal Amplification (LAMP) assay for the early on-site detection of GBS infection from neonatal sepsis blood samples. To develop a LAMP assay, the primers are designed for the selective targeting of a highly conserved segment within the cfb gene encoding the CAMP factor in Streptococcus agalactiae ensuring high specificity.
RESULTS: Rigorous optimization of reaction conditions, including temperature and incubation time, enhances the efficiency of the LAMP assay, enabling rapid and reliable GBS detection within a short timeframe. The diagnostic efficacy of the LAMP assay was evaluated using spiked blood samples by eliminating the DNA extraction step. The simplified colorimetric LAMP assay has the capability to detect S. agalactiae in a neonatal blood sample containing 2 CFU/mL during sepsis. Additionally, the LAMP assay effectively detected S. agalactiae in both the standard and spiked blood samples, with no detectable interference with blood.
CONCLUSIONS: This optimised LAMP assay emerges as a promising tool for early GBS detection, offering a rapid and accurate on-site solution that has the potential to inform timely interventions and improve outcomes in neonatal sepsis cases.
摘要：
背景：新生儿败血症，通常归因于B族链球菌（GBS）感染，对婴儿构成严重的健康风险，要求快速和准确的诊断方法。现有的诊断方法依赖于传统的培养方法，该过程需要大量时间，并且有可能延迟关键的治疗评估。
方法：本研究引入了一种创新的环介导等温扩增（LAMP）检测方法，用于早期现场检测新生儿败血症血样中的GBS感染。为了开发LAMP检测方法,这些引物设计用于选择性靶向无乳链球菌中编码CAMP因子的cfb基因内的高度保守片段,以确保高特异性.
结果：反应条件的严格优化，包括温度和孵育时间，提高了LAMP检测的效率，在短时间内实现快速可靠的GBS检测。通过消除DNA提取步骤，使用加标的血液样品评估了LAMP测定的诊断功效。简化的比色LAMP测定具有检测脓毒症期间含有2CFU/mL的新生儿血液样品中的无乳链球菌的能力。此外,LAMP检测有效地检测了标准和加标血液样本中的无乳链球菌，对血液没有检测到的干扰。
结论：这种优化的LAMP检测方法成为早期GBS检测的有前途的工具，提供快速准确的现场解决方案，有可能及时提供干预措施并改善新生儿败血症病例的结局。
方法：本研究引入了一种创新的环介导等温扩增（LAMP）检测方法，用于早期现场检测新生儿败血症血样中的GBS感染。为了开发LAMP检测方法,这些引物设计用于选择性靶向无乳链球菌中编码CAMP因子的cfb基因内的高度保守片段,以确保高特异性.
结果：反应条件的严格优化，包括温度和孵育时间，提高了LAMP检测的效率，在短时间内实现快速可靠的GBS检测。通过消除DNA提取步骤，使用加标的血液样品评估了LAMP测定的诊断功效。简化的比色LAMP测定具有检测脓毒症期间含有2CFU/mL的新生儿血液样品中的无乳链球菌的能力。此外,LAMP检测有效地检测了标准和加标血液样本中的无乳链球菌，对血液没有检测到的干扰。
结论：这种优化的LAMP检测方法成为早期GBS检测的有前途的工具，提供快速准确的现场解决方案，有可能及时提供干预措施并改善新生儿败血症病例的结局。
