Development, Optimization, and Validation of a Multiplex Real-Time PCR Assay on the BD MAX Platform for Routine Diagnosis of Acanthamoeba Keratitis

Maubon, Danièle; Richarme, Claire; Post, Lucie; Robert, M; Bernheim, Diane; Garnaud, Cécile

doi:10.1016/j.jmoldx.2020.09.001

Cited by 3 publications

(2 citation statements)

References 29 publications

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“…Recently, many nucleic acid amplification tests have been developed that claim improved the diagnosis of AK. [ 17 18 19 ] Molecular methods have an advantage of a shorter turnaround time compared to culture, and the theoretical detection threshold is one ameba per sample. However, the expertise and infrastructure for doing molecular tests are not available at all centers.…”

Section: Discussionmentioning

confidence: 99%

Acanthamoeba keratitis

Ahmed

Rathod

Satpathy

et al. 2022

Tropical Parasitology

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The free-living amebae of genus Acanthamoeba are an important cause of microbial keratitis. The clinical appearance of Acanthamoeba keratitis (AK) usually mimics viral or fungal keratitis. Thus, microbiological workup plays a significant role in the diagnosis and timely treatment of such cases. We report a retrospective case series of seven culture-confirmed AK cases from a tertiary eye care center in North India. Various risk factors and triggers of infection, clinical presentations, microbiological findings, and management of AK are elucidated.

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Section: Discussionmentioning

confidence: 99%

Acanthamoeba keratitis

Ahmed

Rathod

Satpathy

et al. 2022

Tropical Parasitology

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“…These NAAT methods have a high sensitivity and can provide rapid results in contrast to the time-consuming culture procedures. 6 , 7 However, a large diversity of NAATs are in use, and so far, external quality assessment scheme (EQAS) for these tests is lacking. An EQAS involves the comparison of test results of a laboratory to a source outside of that laboratory, allowing an objective assessment of the diagnostic performance of a laboratory.…”

mentioning

confidence: 99%

An International External Quality Assessment Scheme to Assess the Diagnostic Performance of Polymerase Chain Reaction Detection of Acanthamoeba Keratitis

Sarink

Koelewijn

Stelma

et al. 2023

Cornea

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The purpose of this study was to assess the variation in methods and to determine whether an External Quality Assessment Scheme (EQAS) for polymerase chain reaction (PCR) detection of Acanthamoeba keratitis is valuable for the diagnostic process.Methods: A multicenter EQAS was introduced, covering 16 diagnostic laboratories. Using Acanthamoeba castellanii ATCC strain 30010, 3 sets of samples were prepared, containing different amounts of DNA, cysts, or trophozoites. Samples were masked and sent to the participants with instructions for use and a questionnaire concerning the applied methodologies. Special attention in this questionnaire was given to the used pretreatment methods to assess existing variations in these procedures.Results: A large variation in the methodologies and substantial differences in the diagnostic performance were found between participants. In contrast to the DNA samples where all participants had a perfect score, several false negative results were reported for the samples containing cysts or trophozoites. Only 9 participants had an optimal score, whereas one participant reported all samples as negative, one participant reported failures due to inhibition, and the other 5 reported in total 7 false negative results. A clear correlation was noticed between the PCR detection rate and the number of cysts or trophozoites in the sample. Conclusions:The results indicate that a pretreatment procedure can be a risky step in PCR-based detections of Acanthamoeba, but it improves the sensitivity and reliability, especially of samples containing cysts. Therefore, participation in an EQAS is informative for routine diagnostic laboratories and can assist in improving the laboratory procedures used for the diagnosis of Acanthamoeba keratitis.

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