İnfluenza A Virüsünün Tanisinda Hücre Kültürü, Real-time PCR, in-House PCR ve Hizli Test Yöntemlerinin Karşilaştirilmasi

SUMMARY Comparison of Cell Culture, Real-time PCR, in-House PCR and Rapid Tests in Diagnosis of Influenza A Viruses Objective: This study was aimed to evaluate and compare the sensitivity and specificity of cell culture, “reverse transcription real-time polymerase chain reaction” (rRT-PCR), in-house PCR and rapid tests used for the diagnosis of diseases caused by influenza viruses. Materials and Methods: Nasal swabs obtained from a total of 100 patients with Influenza Like Illness (ILI) were tested using three different methods: a) cell culture / immune captureenzyme-linked immunosorbant assay (IC-ELISA) b) RT-PCR and c) rapid test. On the other hand, nasal samples obtained from 50 other patients were also tested by in-house PCR in addition to the three methods mentioned above. In the first group in rRT-PCR primer-probes directed for HA region were used while in the second group in rRT-PCR and in in-house PCR primer-probes directed for M region were used. The samples which yielded conflicting results were further investigated by reverse transcription polymerase chain reaction enzyme hybridization assay (RT-PCR-EHA) using primers targeting different gene regions. Results: In the group in which the three methods were compared, cell culture was considered as the gold standard and the sensitivity of rRT-PCR and rapid tests were determined as 80% and 14,5 and specificities as 91,1% and 100%, respectively. In the group which was composed of 50 samples, positive results in at least two tests were taken as a prerequisite for the true positive results. Accordingly, the sensitivities of cell culture, real time RT-PCR, in-house PCR and rapid tests were 86,7%, 100%, 76,7%, and 10%, and specificities were 100%, 85%, 100% and 100% respectively. Conclusion: As important issues, this study revealed, and emphasized the importance of usage of appropriate primers and probes to increase the sensitivity and specificity of PCR tests, taking quality control measures and when necessary application of different laboratory techniques to reach the true positive results.