[Comparative analysis of LAMP and Real Time PCR methods to detect pathogens of glanders and meliodosis.].

This study compares the effectiveness of two DNA detection methods, LAMP and Real Time PCR, in identifying specific strains of the bacteria Burkholderia mallei and Burkholderia pseudomallei, which cause glanders and melioidosis respectively. The study suggests that Real Time PCR is the more effective method for detecting these pathogens due to issues with sensitivity, specificity, and reliability in the LAMP method.

Comparison of LAMP and Real Time PCR methods

In this study, Loop-mediated Isothermal Amplification (LAMP) and Real Time Polymerase Chain Reaction (PCR) methods were compared for their ability to detect the presence of DNA from two bacteria, Burkholderia mallei and Burkholderia pseudomallei.

• Real Time PCR: In this method, the study showed that the selected primers (short strands of DNA that serve as starting points for DNA synthesis) consistently detected DNA of the specific strains of bacteria they were designed for, and were not confused by DNA of unrelated strains.
• LAMP: By contrast, the LAMP approach proved less reliable. No set of primers showed high sensitivity (the ability to correctly identify positive results) and high specificity (the ability to correctly identify negative results). Furthermore, the LAMP results were less consistent in repeated experiments, which questions the reliability of the method.

Challenges with the LAMP Method

Several reasons were identified that might explain the issues encountered with the LAMP method in this study.

• Complexity of DNA: The genomes of Burkholderia mallei and Burkholderia pseudomallei are rich in GC (guanine and cytosine) regions. Such regions can form strong bonds and create complex secondary structures that are difficult for the LAMP method to handle.
• Unintended DNA synthesis: The LAMP method’s primers were found to direct the synthesis of DNA from genes they were not targeted for, which could lead to false positive results.

Recommendation

Given the difficulties and inconsistencies identified with the LAMP method, the study recommends the use of Real Time PCR for detecting these bacterial strains in complex genetic materials such as the DNA of Burkholderia mallei and Burkholderia pseudomallei. Real Time PCR demonstrated consistent and reliable results and was able to accurately identify the target bacterial DNA.