A Loop-Mediated Isothermal Amplification (LAMP) Assay Specific to Trichomonas tenax Is Suitable for Use at Point-of-Care

Abstract

Trichomonas tenax is a flagellated protozoan that inhabits the human and canine oral cavity in patients with poor oral hygiene and periodontal disease. The loop-mediated isothermal amplification (LAMP) assay could provide clinicians with a quick, cheap and reliable diagnostic test used for the detection of T. tenax in various settings. In this study, we aimed to develop a LAMP assay that can detect T. tenax with high sensitivity and specificity. A set of LAMP primers were specifically designed to detect the ITS and 5.8S rRNA gene of T. tenax. The newly developed LAMP assay was 1000 times more sensitive than conventional PCR. The limit of detection of the LAMP assay was 10 fg of genomic DNA, or 0.2–1 cell. Moreover, the LAMP assay was specific, resulting in no cross-reaction even with a closely related protozoan T. vaginalis or other microorganisms (Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Candida albicans) used. The present LAMP assay can be performed directly without prior DNA extraction, making the assay an easy, fast, cheap, specific and sensitive diagnostic tool for the detection of T. tenax at the point-of-care of both medical and veterinary clinics in developed and developing countries.

Sequence-Specific Electrochemical Genosensor for Rapid Detection of blaOXA-51-like Gene in Acinetobacter baumannii

“…A real-time PCR assay has also been developed for A. baumannii detection [ 56 ], which provides faster results compared with conventional PCR. However, PCR assays require the use of expensive equipment and thus may not be an ideal solution for some countries [ 57 , 58 ]. Accurate and early detection of A. baumannii infections is very essential to efficient healthcare for patients due to the high mortality rate associated with its infections [ 59 ].…”

Recent progress in diagnosis and treatment of Human African Trypanosomiasis has made the elimination of this disease a realistic target by 2030

“…Hence, the LAMP LOD matches the numbers for PCR, but offers the advantage of being performed isothermally. Indeed, this test can be performed at a constant temperature of 58-65 • C. Furthermore, LAMP allows the amplification to be performed using clinical specimens without resulting in inhibition of their enzymes (41,57). In addition, it permits a direct readout either by fully quantitative (and more subjective) techniques such as checking turbidity due to magnesium pyrophosphate precipitate in the reaction tube, or by more qualitative and user-friendly methods such as LFIAs (54,59,60).…”

“…to have the highest amplification products, while 30 min. is sufficient to have a visible signal when a decreased LOD is acceptable ( 56 , 57 ). Reaction rates can be further improved by optimizing concentrations of dNTPs and Mg 2+ ( 54 ).…”

A loop-mediated isothermal amplification-enabled analytical assay for the detection of SARS-CoV-2: A review

“…FIP is the upper inner primer used to initiate the reaction in the first step of template synthesis, and BIP is the lower inner primer used to carry out the second stage of cyclic amplification; the four primers participate in the LAMP reaction process in the initial stage of template synthesis, and only the inner primer is needed in the subsequent cycle reaction process. Under the action of Bst DNA polymerase, nucleic acid amplification is carried out at 60-65°C (Matthew et al, 2022). The detection of the amplified products generally includes three methods: agarose gel electrophoresis, metal ion indicator or dye coloration, and observation of the white magnesium pyrophosphate precipitate, which can be directly observed with the naked eye (Petrusha and Faizuloev, 2020).…”