Detection of fungal contamination by 28S rRNA gene PCR in Ophthalmic viscoelastics
The aim of the work is to evaluate a polymerase chain reaction (PCR) based assay to detect fungal contamination in ophthalmic viscoelastics such as hydroxypropyl methylcellulose and sodium hyaluronate. Viscoelastic samples were artificially contaminated with <10 CFU of Aspergillus brasiliensis (and other fungal contaminants), DNA was extracted and a PCR assay was performed using specific primers for the panfungal genome (28S rRNA). Specificity and sensitivity of the technique were verified. In conclusion, standard methods required 6 to 8 days for fungal contamination detection, whereas PCR based detection was completed within 8 hours. This assay was highly specific to detect fungal contaminants rapidly and no PCR inhibition was observed in ophthalmic viscoelastics.
Fungal contamination of pharmaceutical products can cause not only a serious economic loss to the manufacturer but also cause serious health problems to customers (patients). Over the years, several mould issues associated with pharmaceutical cleanrooms, cold rooms and controlled areas have been reported (1, 2). Additionally, a variety of compounded pharmaceutical products have also lead to more than 200 adverse effects including vision loss, blindness, infection and death (2). The most serious of these effects was a fungal meningitis outbreak in the year 2012. This outbreak was caused by fungal contamination of compounded product steroidal injections and resulted in the death of 64 patients (3). More recently, there have been two similar concurrent outbreaks of vision-threatening endophthalmitis which were reported as 2 environmental moulds contaminating 2 ophthalmic compounded medications labelled as sterile form (4).
If contamination occurs in ophthalmic pharmaceuticals this will lead not only to product loss to manufacturer, but also permanent visual loss to patients. These reported fungal ...