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Download All Background: The genus Acanthamoeba is primarily a free-living protozoan in natural habitats, but also causative agent of human and animal disease. Acanthamoeba serves as host for a variety of pathogenic bacteria such as Mycobacterium avium. Infestation with Acanthamoeba is associated with potentially sight-threatening contact lens-related keratitis, serious infections of other organs and fatal granulomatous amoebic encephalitis. More than 20 species of Acanthamoeba are known, which can be classified into three morphologic 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 been recently associated with human disease. Genotype T4 has been considered the most important genotype in both ocular and CNS infection. According to literature, T4 genotype is the most prevalent Acanthamoeba genotype causing keratitis worldwide (approx. 86%) (Diehl et al., 2021).
Description: ParoReal® Kit Acanthamoeba T4 is is a real-time PCR for the qualitative detection of the DNA (18S rRNA gene) of Acanthamoeba species of genotype T4 (A. castellani, A. lugdunensis, A. mauritaniensis, A. polyphaga, A. rhysodes, A. royreba). T4 genotype is the most prevalent (approx. 86%) Acanthamoeba genotype causing keratitis worldwide. The test does not detect other Acanthamoeba genotypes (T3, T15, T11, and T5) also causing keratitis. Proper specimens are DNA extracts from human clinical specimens associated with keratitis (ocular swabs, corneal biopsies, ocular punctates, corneal scrapings) as well as contact lenses and contact lens solution. Our carefully designed primers and probe ensure highest sensitivity and specificity. The kit consists of an assay mix for the detection of the pathogen as well as a positive control and an internal positive control (IPC) system. To minimize PCR cross-contamination, the included reaction mix contains dUTP and uracil-N glycosylase (UNG).
| Order Number | Reactions | Channel Pathogen | Channel IPC | Target |
|---|---|---|---|---|
| DHUP00153 | 50 | FAM | Cy5 | 18S rRNA |
Shipping Temperature: +4 °C cool packs
- Amplification and detection: 18S rRNA gene of Acanthamoeba genotype T4
- Real-time PCR with rapid hot-start Taq DNA polymerase
- ROX™ dye as passive reference
- Internal Positive Control System to exclude false-negative results
- Optimized to handle PCR inhibitors
- PCR platforms: runs on all established standard real-time PCR platforms
- Harmonized thermal profiles to run RNA and DNA samples simultaneously
PCR platforms: ParoReal® Kit Acanthamoeba T4 was validated with the ABI® 7500 Fast Real-Time PCR System (fast cycle parameters are not supported, Thermo Fisher Scientific), and was also tested with Mx3005P® (Agilent), qTOWER3G (Analytik Jena), MIC instrument (bio molecular systems), LightCycler® 480 II (Roche), CFX Opus Real-time PCR System (Bio-Rad), CFX Dx (Bio-Rad), T-COR 8 (Tetracore) and cobas Z 480 Analyzer (Roche). It is also compatible with other real-time PCR instruments which detect and differentiate fluorescence in the FAM and Cy5 channel (e.g. QuantStudio™ 5 / 6 / 7 (Thermo Fisher Scientific)).