title: Detection and Typing of Human Papilloma Viruses by Nested Multiplex Polymerase Chain Reaction Assay in Cervical Cancer.
abstract: BACKGROUND
  Cervical cancer is the leading cause of death from cancer in under-developed countries. Human papilloma virus (HPV) 16 and 18 are the most prevalent types associated with carcinogenesis in the cervix. Conventional Polymerase Chain Reaction (PCR), type-specific and consensus primer-based PCR followed by sequencing, Restriction Fragment Length Polymorphism (RFLP) or hybridization by specific probes are common methods for HPV detection and typing. In addition, some researchers have developed a multiplex PCR for simultaneous detection and typing of different HPVs.
  
  
  OBJECTIVE
  The aim of the present study was to investigate the prevalence of HPV infection and its types in cervical Squamous Cell Carcinoma (SCC) using the Nested Multiplex PCR (NMPCR) assay.
  
  
  METHODS
  Sixty-six samples with histologically confirmed SCC were evaluated. Total DNA was isolated by phenol-chloroform extraction and ethanol precipitation. Nested multiplex PCR was performed with first-round PCR by GP-E6/E7 consensus primers for amplification of the genomic DNA of all known mucosal HPV genotypes and second-round PCR by type-specific multiplex PCR primer cocktails.
  
  
  RESULTS
  Human papilloma virus infection was detected in 78.8% of samples, with the highest prevalence of HPV 16 (60.6%) while concurrent infections with two types was detected in 10.6%.
  
  
  CONCLUSIONS
  The NMPCR assay is more convenient and easy for analysis of results, which is important for fast diagnosis and patient management, in a type-specific manner.
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Seyed
authors: S
authors: Jalal Kiani
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Somayeh
authors: S
authors: Shatizadeh Malekshahi
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Zohreh
authors: Z
authors: Yousefi Ghalejoogh
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Nastaran
authors: N
authors: Ghavvami
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Nazanin Zahra
authors: NZ
authors: Shafiei Jandaghi
authors: Pathology Department, School of Medicine, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Reza
authors: R
authors: Shahsiah
authors: Pathology Department, School of Medicine, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Isa
authors: I
authors: Jahanzad
authors: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors: Jila
authors: J
authors: Yavarian
keywords: Carcinoma; Human Papillomavirus 16; Multiplex Polymerase Chain Reaction; Squamous Cell