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

Filippis, Ivano de; Azevedo, Aline Carvalho de; Lima, Irene de Oliveira; Ramos, Nicolle F Lima; Andrade, Cláudia; Almeida, Antonio Ecc de

doi:10.2144/btn-2022-0089

Cited by 2 publications

(3 citation statements)

References 20 publications

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“…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].…”

Section: Discussionmentioning

confidence: 99%

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

Jarzyniak,

Narożna

2024

Agronomy

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Soybean [Glycine max (L.) Merr.] is one of the most important and oldest crops. Due to its ability to form symbiotic interactions with nitrogen-fixing bacteria, it is a valuable source of nitrogen for agriculture and proteins for humans and livestock. In Europe, for instance, in Poland, the soybean cultivation area is still not large but is gradually increasing due to climate change. The lack of indigenous soybean microsymbionts in Polish soils forces the application of commercial strains to establish effective symbioses. Fast and reliable identification methods are needed to study the persistence, competitiveness, and dispersal of bradyrhizobia introduced as inocula. Our study aimed to apply real-time PCR coupled with high-resolution melting curve (HRM) analysis to detect and differentiate bacterial strains occupying soybean nodules. HRM-PCR was performed on crude extracts from nodules using primers specific for recA, a highly conserved nonsymbiotic gene. By comparing them with the reference strains, we were able to identify and assign Bradyrhiobium strains that had been introduced into field locations in Poland. In conclusion, HRM analysis was proven to be a fast and accurate method for identifying soybean microsymbionts and might be successfully used for identifying other legume-nodulating bacteria.

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Section: Discussionmentioning

confidence: 99%

“…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.…”

Section: Discussionmentioning

confidence: 99%

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

Jarzyniak,

Narożna

2024

Agronomy

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Sánchez-Paz,

Encinas-García,

Mendoza-Cano

2023

Fishes

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In this study, the primers of different protocols for the detection of White Spot Syndrome Virus (WSSV) were analyzed in silico to evaluate their potential performance in PCR. As with any biological entity, this virus evolves constantly. Thus, this analysis showed that a few primers, including those recommended by the World Organization for Animal Health (WOAH), might mismatch with some isolates of WSSV, specially with isolates more recently sequenced. Furthermore, a set of primers recommended by WOAH, showed the potential to self-dimer and form hairpin loop structures, which could affect the efficiency of PCR, resulting in an inaccurate diagnostic result. From our perspective, and considering the evolutionary trajectory of this virus, it may be time for the WOAH to update the PCR protocols recommended for WSSV detection, which remains as a highly prevalent and lethal virus.

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

Cited by 2 publications

References 20 publications

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

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

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

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