Specific detection of common pathogens of acute bacterial meningitis using an internally controlled tetraplex-PCR assay

Farahani, Hamidreza; Ghaznavi‐Rad, Ehsanollah; Mondanizadeh, Mahdieh; Samiee, Siamak Mirab; Khansarinejad, Behzad

doi:10.1016/j.mcp.2016.07.002

Cited by 2 publications

(2 citation statements)

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“…This confirmed the utility of SPNA45_01710 and NMO_1242 genes as effective targets for S. pneumoniae and N. meningitidis , respectively. Various biomarkers for S. pneumoniae ( ply , lytA and Spn9802) and N. meningitidis ( ctrA , sodC and siaD ) have been used for PCR and LAMP assay development [ 6 , 15 , 18 , 19 , 28 , 29 , 30 , 31 ]; however, many of these targets have been identified as inefficient [ 28 , 29 , 32 ]. We propose that the SPNA45_01710 gene, and the sequence between bases 450–1800 of the NMO_1242 gene, are potential novel regions-of-interest for the identification of S. pneumoniae and N. meningitidis , respectively.…”

Section: Discussionmentioning

confidence: 99%

“…Streptococcus pneumoniae , Neisseria meningitidis and Haemophilus influenzae are leading etiological agents of bacterial meningitis infection, with S. pneumoniae and N. meningitidis responsible for over 80% of all incidents in regions with routine H. influenzae vaccination [ 5 , 6 , 7 ]. Despite this, H. influenzae related meningitis remains a serious threat, especially among children under 5 years of age in regions without routine immunization [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of an Internally Controlled Multiplex Tth Endonuclease Cleavage Loop-Mediated Isothermal Amplification (TEC-LAMP) Assay for the Detection of Bacterial Meningitis Pathogens

Higgins

Clancy

Cormican

et al. 2018

IJMS

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Bacterial meningitis infection is a leading global health concern for which rapid and accurate diagnosis is essential to reduce associated morbidity and mortality. Loop-mediated isothermal amplification (LAMP) offers an effective low-cost diagnostic approach; however, multiplex LAMP is difficult to achieve, limiting its application. We have developed novel real-time multiplex LAMP technology, TEC-LAMP, using Tth endonuclease IV and a unique LAMP primer/probe. This study evaluates the analytical specificity, limit of detection (LOD) and clinical application of an internally controlled multiplex TEC-LAMP assay for detection of leading bacterial meningitis pathogens: Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae. Analytical specificities were established by testing 168 bacterial strains, and LODs were determined using Probit analysis. The TEC-LAMP assay was 100% specific, with LODs for S. pneumoniae, N. meningitidis and H. influenzae of 39.5, 17.3 and 25.9 genome copies per reaction, respectively. Clinical performance was evaluated by testing 65 archived PCR-positive samples. Compared to singleplex real-time PCR, the multiplex TEC-LAMP assay demonstrated diagnostic sensitivity and specificity of 92.3% and 100%, respectively. This is the first report of a single-tube internally controlled multiplex LAMP assay for bacterial meningitis pathogen detection, and the first report of Tth endonuclease IV incorporation into nucleic acid amplification diagnostic technology.

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