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. 2015 Dec 26;8(12):e26441.
doi: 10.5812/jjm.26441. eCollection 2015 Dec.

Detection and Typing of Human Papilloma Viruses by Nested Multiplex Polymerase Chain Reaction Assay in Cervical Cancer

Seyed Jalal Kiani  1 , Somayeh Shatizadeh Malekshahi  1 , Zohreh Yousefi Ghalejoogh  1 , Nastaran Ghavvami  1 , Nazanin Zahra Shafiei Jandaghi  1 , Reza Shahsiah  2 , Isa Jahanzad  2 , Jila Yavarian  1
Affiliations

Detection and Typing of Human Papilloma Viruses by Nested Multiplex Polymerase Chain Reaction Assay in Cervical Cancer

Seyed Jalal Kiani et al. Jundishapur J Microbiol. .

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.

Objectives: 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.

Patients and 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.

Keywords: Carcinoma; Human Papillomavirus 16; Multiplex Polymerase Chain Reaction; Squamous Cell.

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Figures

Figure 1.
Figure 1.. Beta-Globin Gene Amplification
A 110-bp product of the human beta-globin housekeeping gene was amplified as an internal control.
Figure 2.
Figure 2.. Agarose Gel Electrophoresis for Nested Multiplex Polymerase Chain Reaction Products of Cocktail-I
The most prevalent types of HPV in cervical cancer samples were high-risk HPV types 16 and 18. Concurrent infection was detected in seven samples (10.6%).
Figure 3.
Figure 3.. Human Papilloma Virus Typing by the Nested Multiplex Polymerase Chain Reaction Assay
The most prevalent types of HPV in cervical cancer samples were high-risk HPV types 16 and 18. Concurrent infection was detected in seven samples (10.6%).
See this image and copyright information in PMC

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