Banerjee Shukla, K Mithun H, Shastry Rajesh P
Division of Microbiology and Biotechnology, Yenepoya Research Centre, Yenepoya (Deemed to be University), University Road, Deralakatte, Mangalore, 575018, India.
Department of Pediatrics, Yenepoya Medical College and Hospital, Yenepoya (Deemed to be University), Deralakatte, Mangalore, 575018, India.
Mol Biol Rep. 2024 Jul 13;51(1):811. doi: 10.1007/s11033-024-09705-0.
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.
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.
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.
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.
新生儿败血症通常归因于B族链球菌（GBS）感染，对婴儿构成重大健康风险，需要快速准确的诊断方法。现有的诊断方法依赖于传统培养方法，这一过程需要大量时间，并且有可能延迟关键的治疗评估。
本研究引入了一种创新的环介导等温扩增（LAMP）检测法，用于从新生儿败血症血样中早期现场检测GBS感染。为了开发LAMP检测法，设计引物以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。
对反应条件（包括温度和孵育时间）进行严格优化，提高了LAMP检测法的效率，能够在短时间内快速可靠地检测GBS。通过省去DNA提取步骤，使用加标血样评估了LAMP检测法的诊断效力。简化的比色LAMP检测法能够在败血症期间检测含有2 CFU/mL的新生儿血样中的无乳链球菌。此外，LAMP检测法在标准血样和加标血样中均有效检测到无乳链球菌，且对血液无明显干扰。
这种优化的LAMP检测法成为早期检测GBS的一种有前景的工具，提供了一种快速准确的现场解决方案，有可能为新生儿败血症病例的及时干预提供依据并改善治疗结果。
