PCR-based approach for qualitative molecular analysis of six neurotropic pathogens

R. FERESE, L. SCORZOLINI, R. CAMPOPIANO, V. ALBANO, A. M. GRIGUOLI, E. GIARDINA, S. SCALA, L. RYSKALIN, C. D'ALESSIO, S. ZAMPATTI, R. FANTOZZI, M. STORTO (+2 others)
2017 Acta virologica  
In the last few years, polymerase chain reaction analysis is frequently required to improve the detection of pathogen infections in central nervous system as a potential cause of neurological disorders and neuropsychiatric symptoms. The goal of this paper is to set up a fast, cheap and reliable molecular approach for qualitative detection of six neurotropic pathogens. A method based on PCR has been designed and implemented to guarantee the qualitative DNA detection of herpes simplex virus types
more » ... 1 and 2 (HSVI/II), Epstein-Barr virus (EBV), cytomegalovirus (CMV), varicella-zoster virus (VZV), rubella virus (RUBV) and Toxoplasma gondii in the cerebrospinal fluid, where otherwise they are barely detectable. Each PCR assay was tested using dilutions of positive controls, which demonstrated a sensitivity allowing to detect up to 10 2 copies/ml in PCR and 10 copies/ ml in real-time PCR for each pathogen. Once been set up, the protocol was applied to evaluate the cerebrospinal fluid from 100 patients with suspected infectious diseases of the central nervous system and 50 patients without any infection. The method allowed to identify 17 positive cerebrospinal fluid with polymerase chain reaction and 22 with real-time PCR (RT-PCR), respectively. Therefore, application of RT PCR allows a fast and sensitive evaluation of neurotropic DNA pathogens in the course of diagnostic routine within neurological units. nervous system; CSF = cerebrospinal fluid; EBV = Epstein-Barr virus; HSV I/II = herpes simplex virus types 1 and 2; HV = herpes virus; JC = John Cunningham; RT-PCR = real-time PCR; RUBV = rubella virus; TGR = Toxoplasma gondii; VZV = varicella-zoster virus
doi:10.4149/av_2017_305 pmid:28854791 fatcat:lxgjymtjzna4ta3k3lqlgvlca4