A DAMP-Based Assay for Rapid and Affordable Diagnosis of Bacterial Meningitis Agents: Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae

被引:2
作者
Shkodenko, Liubov A. [1 ]
Mohamed, Al-Abbass [1 ]
Ateiah, Muhannad [1 ]
Rubel, Maria S. [1 ]
Koshel, Elena I. [1 ]
机构
[1] ITMO Univ, Lab DNA Nanosensor Diagnost, Lomonosova St 9, St Petersburg 191002, Russia
基金
俄罗斯科学基金会;
关键词
isotheral amplification; DAMP; CNS infections; meningitis; DNA diagnostics; POC; diagnostic assay; COMMUNITY-ACQUIRED BACTERIAL; NUCLEIC-ACID AMPLIFICATION; URACIL-DNA-GLYCOSYLASE; ISOTHERMAL AMPLIFICATION; CARE DIAGNOSTICS; VISUAL DETECTION; PCR ASSAY; POINT; TECHNOLOGIES;
D O I
10.3390/ijms25158282
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The rapid and accurate diagnosis of meningitis is critical for preventing severe complications and fatalities. This study addresses the need for accessible diagnostics in the absence of specialized equipment by developing a novel diagnostic assay. The assay utilizes dual-priming isothermal amplification (DAMP) with unique internal primers to significantly reduce non-specificity. For fluorescence detection, the dye was selected among Brilliant Green, Thioflavin T, and dsGreen. Brilliant Green is preferred for this assay due to its availability, high fluorescence level, and optimal sample-to-background (S/B) ratio. The assay was developed for the detection of the primary causative agents of meningitis (Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae), and tested on clinical samples. The developed method demonstrated high specificity, no false positives, sensitivity comparable to that of loop-mediated isothermal amplification (LAMP), and a high S/B ratio. This versatile assay can be utilized as a standalone test or an integrated assay into point-of-care systems for rapid and reliable pathogen detection.
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页数:12
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共 64 条
  • [61] Investigation and validation of labelling loop mediated isothermal amplification (LAMP) products with different nucleotide modifications for various downstream analysis
    Warmt, Christian
    Yaslanmaz, Ceren
    Henkel, Joerg
    [J]. SCIENTIFIC REPORTS, 2022, 12 (01)
  • [62] A systematic review of the diagnostic accuracy of Loop-mediated-isothermal AMPlification (LAMP) in thediagnosis of invasive meningococcal disease in children
    Waterfield, Thomas
    Fairley, Derek
    Blackwood, Bronagh
    McKenna, James
    Shields, Michael D.
    [J]. BMC PEDIATRICS, 2019, 19 (1)
  • [63] INSIGHT: A population-scale COVID-19 testing strategy combining point-of-care diagnosis with centralized high-throughput sequencing
    Wu, Qianxin
    Suo, Chenqu
    Brown, Tom
    Wang, Tengyao
    Teichmann, Sarah A.
    Bassett, Andrew R.
    [J]. SCIENCE ADVANCES, 2021, 7 (07):
  • [64] Meningitis in adults: diagnosis and management
    Young, Nicholas
    Thomas, Mark
    [J]. INTERNAL MEDICINE JOURNAL, 2018, 48 (11) : 1294 - 1307