Accurate, Fast and Cost-Effective Simultaneous Detection of Bacterial Meningitis by Qualitative PCR with High-Resolution Melting

Abstract

Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are leading causes of meningitis and acute invasive infections. PCR-based methods are widely used for the diagnosis and surveillance of bacterial pathogens because of their high sensitivity, specificity and high-throughput capabilities compared with conventional laboratory methods. This study evaluated a high-resolution melting qualitative PCR analysis method for the simultaneous detection of these three pathogens. The assay has been optimized to detect three species-specific genes of each organism isolated from clinical samples, enabling accurate identification of the etiological agent. The method proved to be highly sensitive and cheaper than the real-time PCR TaqMan® system because it is probe-free; it could be used for the diagnosis of invasive diseases in public health laboratories of developing countries.

Rapid Identification of Rhizobia Nodulating Soybean by a High-Resolution Melting Analysis

“…To date, this technique has been mainly applied to evaluate microbial diversity in clinical research, diagnostics, and food science [21][22][23][24][25][26][27]. By using HRM-PCR, researchers were able to detect bacterial meningitis [34], diagnose drug-resistant tuberculosis [26], identify lactic acid bacteria [23,25], and determine seafood quality [24].…”

“…The verification of HRM results through sequencing has been conducted in several other studies [23,24,53]. Additionally, alternative species-specific PCRs [25,34] or commercially available tests [44] have also been used to validate the method.…”

In Silico Evaluation of the PCR Performance of Different Tests for Detection of WSSV

“…PCR-based protocols are arguably the most widely used molecular methods for diagnosing and surveilling pathogens because of their high sensitivity, specificity, and highthroughput capabilities [35]. However, a suitable PCR protocol depends on numerous factors in the amplification reaction that may affect its efficiency.…”