Establishment of a Rapid LAMP Assay for Aeromonas hydrophila and Comparison with the Application of qPCR

Gao, Zihui; Yang, Chunhua; Zhang, Xiaobo; Hu, Bing; Zhang, Huang; Kuang, Wendong; Zheng, Qian; Cao, Jijuan

doi:10.3390/metabo13070841

Cited by 2 publications

(1 citation statement)

References 23 publications

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“…The study also explored the use of crude nucleic acid extract as compared to pure genomic DNA and found a sensitivity of 559 pg with the crude extract, indicating a rapid detection level of actual samples. [ 50 ] In another study, LAMP and qPCR were developed for the diagnosis of the herpes B virus infection. The LAMP and qPCR detected 50 and 10 copies of the target fragment, respectively.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of loop-mediated isothermal amplification method for efficient detection of the periodontopathic bacteria Porphyromonas gingivalis

Sangolli,

Kugaji,

Ray

et al. 2024

Journal of Indian Society of Periodontology

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Background: Periodontitis is a multifactorial, polymicrobial oral inflammatory illness brought on by oral pathogens. Porphyromonas gingivalis is a Gram-negative, obligatory anaerobic, black-pigmented coccobacillus and is regarded as a primary etiological factor in the progression of periodontitis. Rapid, highly senstitive and specific detection methods are emerging. The present study aimed to evaluate the loop-mediated isothermal amplification (LAMP) technique for efficiently detecting P. gingivalis from subgingival plaque samples of chronic periodontitis patients. Materials and Methods: This study included 50 subgingival plaque samples from patients suffering from chronic periodontitis. The DNA (Deoxyribonucleic acid) was extracted by the “modified proteinase K” method. A set of six primers, targeting the pepO gene of P. gingivalis, was used for conducting LAMP. The amplification was visualized by naked-eye detection and agarose electrophoresis. Conventional polymerase chain reaction (PCR) and real-time qantitative PCR (qPCR) were carried out by targeting the 16SrRNA (16S ribosomal ribonucleic acid) gene of P. gingivalis. Results: The results showed that LAMP detected P. gingivalis in 40 out of 50 samples (80%). Whereas, qPCR and conventional PCR technique detected P. gingivalis in 38 (76%) and 33 (66%) samples respectively. The sensitivity and specificity of the LAMP method were 94.87% and 90.90%, respectively. With qPCR, the sensitivity and specificity were found to be 92.30% and 81.81%, respectively, whereas, with conventional PCR, it was found to be 76.92% and 72.72%, respectively. Conclusion: LAMP is an efficient technique for quick, accurate, and reliable identification of P. gingivalis from subgingival plaque samples. The technique needs to be validated analytically, and further studies can be conducted by taking saliva and/or gingival crevicular fluid samples from periodontitis patients.

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Section: Discussionmentioning

confidence: 99%

Evaluation of loop-mediated isothermal amplification method for efficient detection of the periodontopathic bacteria Porphyromonas gingivalis

Sangolli,

Kugaji,

Ray

et al. 2024

Journal of Indian Society of Periodontology

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Field-Applicable Loop-Mediated Isothermal Amplification for the Detection of Seven Common Human Papillomavirus Subtypes

Li,

Tan,

et al. 2024

TropicalMed

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Persistent HPV infection is a major risk factor for the subsequent development of cervical cancer. LAMP is simple and suitable for field detection in the resource-limited settings. In this study, hydroxy naphthol blue (HNB)-based visual LAMP and evagreen-based fluorescent LAMP coupled with a microfluidic chip (LAMP-chip) were established for the field detection of seven subtypes of HPV. The analytical sensitivity was 19–233 copies/reaction. The overall clinical sensitivity was 97.35% for visual LAMP and 98.23% for LAMP-chip. Both LAMP assays exhibited 100% specificity and were completed in less than 50 min. Additionally, both assays did not require complicated nucleic acid extraction and purification steps. A complete quality control monitoring system (including internal control, positive quality control and negative control) in the LAMP assays further ensured the credibility of the results. Our findings demonstrated that the proposed LAMP assays have the potential to be applied in the testing of common HPV DNA in field investigations (visual LAMP) or within communities and primary health centers (LAMP-chip).

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Establishment of a Rapid LAMP Assay for Aeromonas hydrophila and Comparison with the Application of qPCR

Cited by 2 publications

References 23 publications

Evaluation of loop-mediated isothermal amplification method for efficient detection of the periodontopathic bacteria Porphyromonas gingivalis

Evaluation of loop-mediated isothermal amplification method for efficient detection of the periodontopathic bacteria Porphyromonas gingivalis

Field-Applicable Loop-Mediated Isothermal Amplification for the Detection of Seven Common Human Papillomavirus Subtypes

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