- Real-Time PCR assay for Acanthamoeba detection and internal positive DNA control (both amplification and extraction of parasite DNA).
- Target for internal DNA positive control
- DNA reaction mixture
- Nuclease-free water
- Acanthamoeba positive control
The ParoReal Acanthamoeba T4 kit is a CE IVD marked real-time PCR assay for quantitative detection of DNA (18S rRNA gene) of Acanthamoeba genotype T4 species (A. castellani, A. lugdunensis, A. mauritaniensis, A. polyphaga, A. rhysodes, A. royreba). Appropriate samples are DNA extracts from human clinical specimens associated with ocular keratitis (eye swabs, corneal biopsies, corneal scrapings) as well as contact lenses and contact lens solution.
Acanthamoeba is a free-living protozoan in natural habitats, but it is also an etiologic agent in humans and animals.
Acanthamoeba infestation is associated with contact lens keratitis, which in addition to being a visual hazard, can lead to severe infections of other organs and the usually fatal granulomatous amoebic encephalitis.
There are more than 20 known species of Acanthamoeba that are classified into three morphological groups:(Group I, II and III) and 22 genotypes (T1-T22).
Some species: (A. castellanii (T4), A. lugdunensis (T4), A. polyphaga (T4), A. rhysodes (T4), A. quina (T4), A. palestinensis (T2), A. griffinii (T3), A. lenticulata (T5), A. astronyxis (T7), A. culbertsoni (T10), A. hatchetti (T11), A. healyii (T12), A. byersi (T18), A. divionensis) have recently been associated with disease. The T4 genotype has been considered the most important genotype in both ocular and central nervous system infections. According to the literature, T4 genotype is the most common genotype of Acanthamoeba causing keratitis worldwide (about 86%) (Diehl et al., 2021).
How to best combat Acanthamoeba keratitis
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The sensitivity of the culture method ranges between 33% and 67%, and the final results of the cultures may take more than a week. In a pathology where any delay in treatment can lead to worse outcomes. The ParoReal® Kit Acanthamoeba T4 increases sensitivity up to 88% and reduces diagnostic time.
Omar Shareef, Sana Shareef, and Hajirah N. Saeed (2023), New Frontiers in Acanthamoeba Keratitis Diagnosis and Management. Biology, 10.3390/biology12121489.
The effectiveness of IVCM can vary greatly depending on the operator. Furthermore, the region and corneal area that IVCM can capture is limited, which may result in scans of regions not affected by AK. In cases of stromal inflammation, false negatives can occur due to the masking of Acanthamoeba cysts by inflammatory cells and edema. Conversely, false positives may occur if macrophages are incorrectly identified as Acanthamoeba cysts.
ParoReal® Kit Acanthamoeba T4 eliminates operator-dependent variability, allowing for standardization in the analysis and interpretation of results, reducing the risk of false negatives and false positives. It is a method accessible to all laboratories, even those without the specific microbiological expertise required for identifying Acanthamoeba.
Omar Shareef, Sana Shareef, and Hajirah N. Saeed (2023), New Frontiers in Acanthamoeba Keratitis Diagnosis and Management. Biology, 10.3390/biology12121489.
For the Speed of Results: Less than 90 minutes, while the culture method (with a sensitivity ranging from 33% to 67%) can take more than a week. Prompt diagnosis of Acanthamoeba keratitis is crucial due to the severity of the disease and the intense pain associated with it.
For its Effectiveness: It standardizes the diagnosis of Acanthamoeba and enhances diagnostic capability compared to microscopy and culture examination.
For its Ease of Use: It detects Acanthamoeba T4 DNA (18S rRNA gene) from direct samples (corneal scraping, contact lenses and storage fluid, and ocular swab).
Because the Test Can Be Performed Alongside All Other Tests in the ViroReal®, BactoReal®, and MycoReal® lines and is compatible with most laboratory instruments.
For its Validation: ParoReal® Acanthamoeba T4 is the only Real-Time PCR test that is CE IVD validated.