Detection of chronic toxoplasmosis in the brain of mice using loop-mediated isothermal amplification (LAMP) and conventional PCR

Hegazy, Mona K.; Saleh, Nora E.; Aboukamar, Wafaa A.

doi:10.1016/j.exppara.2023.108556

Cited by 2 publications

(2 citation statements)

References 67 publications

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“…Various molecular techniques for detecting T. gondii in clinical samples represented a significant advance in the diagnosis of toxoplasmosis [ 14 , 15 ]. In immunocompromised patients, acute toxoplasmosis, including toxoplasma encephalitis and disseminated toxoplasmosis, is fatal.…”

Section: Discussionmentioning

confidence: 99%

Development of a LAMP method with lateral flow DNA chromatography to diagnose toxoplasmosis in immunocompromised patients

Mikita,

Mori,

Komine

et al. 2024

Trop Med Health

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Background Rapid and accurate diagnosis of toxoplasmosis is critical, particularly for immunocompromised patients. Several molecular methods could have value for toxoplasmosis diagnosis, but often require sophisticated and expensive equipment, and as such are impractical for use in resource-limited countries. Our study aimed to develop a new rapid diagnostic test for toxoplasmosis that can be used in developed countries as well as low- or middle-income countries. Methods Common primers for conventional loop-mediated isothermal amplification (LAMP) and the new LAMP DNA chromatography method were designed based on a 529-bp repeat present in Toxoplasma gondii genomic DNA. A total of 91 clinical samples from 44 patients suspected of having toxoplasmosis who were treated at several hospitals across Japan were tested using the new LAMP DNA chromatography method, conventional LAMP, and nested PCR and the sensitivity and specificity of the methods was compared. Results The LAMP DNA chromatography method showed better sensitivity and specificity (68.2% and 100%, respectively) compared with the nested PCR (45.4% and 100%, respectively) and conventional LAMP (63.6% and 100%, respectively) methods for diagnosis of toxoplasmosis in immunocompromised patients. LAMP DNA chromatography also has better sensitivity and specificity (75% and 100%, respectively) than nested PCR (50.0% and 93.5%, respectively) and conventional LAMP (62.5% and 100%, respectively) to diagnose toxoplasma encephalitis using CSF samples. Conclusion We developed a LAMP DNA chromatography method to detect T. gondii DNA in clinical samples. This method also successfully detected T. gondii DNA in CSF from patients with toxoplasma encephalitis. This newly developed method can be a valuable rapid diagnostic test for toxoplasmosis in a range of settings, including resource-limited areas like those in low- or middle-income countries.

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Section: Discussionmentioning

confidence: 99%

Development of a LAMP method with lateral flow DNA chromatography to diagnose toxoplasmosis in immunocompromised patients

Mikita,

Mori,

Komine

et al. 2024

Trop Med Health

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“…Similarly, immunohistochemistry (IHC) and immunofluorescence assay (IF) have been applied to detect T. gondii ME49 infection [ 5 ]. Moreover, the polymerase chain reaction (PCR) has been continually optimized as a promising detection method for these infections [ 6 , 7 ]. The use of the B1 gene and the ITS-1 region as primers in PCR was reported to be highly sensitive in quantifying T. gondii ME49 infections [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques

Kang,

Mao,

Kim

et al. 2023

Parasites Hosts Dis

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Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

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Detection of chronic toxoplasmosis in the brain of mice using loop-mediated isothermal amplification (LAMP) and conventional PCR

Cited by 2 publications

References 67 publications

Development of a LAMP method with lateral flow DNA chromatography to diagnose toxoplasmosis in immunocompromised patients

Development of a LAMP method with lateral flow DNA chromatography to diagnose toxoplasmosis in immunocompromised patients

The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques

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