A simple and rapid approach for human herpesvirus type 8 subtype characterization using single base extension

Sequence analysis of the ORFK1 human herpesvirus type 8 (HHV-8) allows identification six major subtypes (A–F), which are related to migrations and clinical progression Kaposi's sarcoma. Sequencing subsequent phylogenetic is considered be most reliable method for HHV-8 genotyping. However, it exhibits challenges limitations. Herein, we designed validated a single base extension (SBE) protocol characterization subtypes. A nested polymerase chain reaction (PCR) was carried out amplify small 294-bp PCR product encompassing four nucleotide polymorphisms at positions 360, 406, 465 527 genome. Finally, multiplex SBE technique developed in 20 samples previously genotyped by analysis. The patterns obtained this could successfully discriminate between typing results completely matched with those ‘gold standard’ all analysed samples. This can reliably identify A, B C, prevalent ones worldwide, remaining (D, E F). useful as an efficient, rapid low-cost screening viral genotyping tube, particularly low-quality DNA, easy data interpretation.