Additional comments

Failure to detect the presence of certain viral types can have a significant impact on patient management. For example, in transplant patients early detection of BK virus (BKV) reactivation is important as this is often the precursor to BKV associated disease, including hemorrhagic cystitis. Notably JCV and BKV are often shed simultaneously in human urine. Therefore, the presence of JCV strains or otherwise sequence polymorphism of the target binding sites could confound the detection of BKV in the urine of transplant patients, reducing the potential to predict the onset of disease.

Co-infections of HSV types 1 and 2 are less common and the failure to detect one of these viruses is arguably of little importance to clinical management and treatment. Nonetheless, incorrect typing of HSV can have significant psychological and social implications, even though the clinical management of the patient is unaffected.

These issues highlight the limitations of hybridization probes used in melting curve analysis for viral characterization in a clinical setting. If the accurate identification of a specific virus type carries clinical importance then careful consideration must be given to the detection methods used in the assay. Preferably, such formats should not rely on consensus oligonu-cleotide sequences but use type-specific primers and probes.

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