The detection of Simian virus 40 in mesotheliomas from New Zealand and England using real time FRET probe PCR protocols.

نویسندگان

  • F Mayall
  • K Barratt
  • J Shanks
چکیده

AIMS To detect Simian virus 40 (SV40) DNA in mesotheliomas from New Zealand and from England using novel real time FRET probe polymerase chain reaction (PCR) protocols. METHODS Twenty four mesotheliomas from New Zealand (Central North Island) and 32 mesotheliomas from England (Greater Manchester region) were examined. Two real time FRET probe PCR protocols were optimised and their analytical sensitivity compared using dilutions of SV40 DNA. A conventional SV40 large tumour antigen protocol with detection by probe hybridisation and chemiluminescent Southern blotting was also optimised. RESULTS Both real time PCR protocols had the same analytical sensitivity, detecting down to 10(-6) pg of SV40 DNA for each reaction, approximately one SV40 copy. All of the 56 mesothelioma samples contained amplifiable beta globin DNA, but none contained amplifiable SV40 DNA with the conventional large T antigen PCR-Southern blotting protocol, or the two real time FRET probe PCR protocols. The positive and negative controls gave the expected results. There was no evidence of inhibition. CONCLUSIONS There is abundant evidence in the literature for the presence of SV40 in mesotheliomas. However, this study found no evidence of SV40 in mesotheliomas from England and New Zealand. The extensive use of SV40 contaminated polio vaccine in New Zealand does not seem to have resulted in SV40 associated mesotheliomas.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Introducing a New SYBR green Real-time PCR for Detection of SARS-CoV2 Virus Genome

Background and purpose: There are various methods for molecular detection of SARS-CoV2 genome among which, PCR-based methods are the most reliable for making diagnosis. The majority of approved PCR kits for detection of Coronavirus are based on TaqMan real-time PCR which is expensive due to incorporating fluorescent and quencher-harboring probe. The aim of this study was to design a simple and ...

متن کامل

Development and Evaluation of Real-Time RT-PCR Test for Quantitative and Qualitative Recognition of Current H9N2 Subtype Avian Influenza Viruses in Iran

Avian influenza H9N2 subtype viruses have had a great impact on Iranian industrial poultry production economy since introduction in the country. To approach Rapid and precise identification of this viruses as control measures in poultry industry, a real time probe base assay was developed to directly detect a specific influenza virus of H9N2 subtype -instead of general detection of Influenza A ...

متن کامل

Evaluation of a new set of Real-Time PCR for Brucella detection within human and animal samples

A quantitative TaqMn Real-Time PCR assay was developed and its diagnostic value on human serum and livestock samples were evaluated. Brucella species could be distributed through communities as a biological agent. Rapid detection of biological threat agents is critical for timely therapeutic administration. Quantitative real-time PCR provides a rapid, sensitive and specific tool for molecular i...

متن کامل

Rapid and accurate diagnosis of Foot-and-mouth disease virus by Real-time PCR in field samples

During 2010-2011, Real-time PCR procedure was used to detecting FMDV RNA on 147 epithelium samples from the field. In this survey, for Real-time PCR from 3D gene segment as conserve region selected for tracking all of seven serotypes FMDV. The assay detected the viral RNA in all serotypes of FMDV. The rRT-PCR specifically detected FMD virus in sample with greater sensitivity than our convention...

متن کامل

Appling real time RT-PCR for bluetongue virus detection in Iran

During 2009-10, real time RT-PCR and conventional RT-PCR techniques Used for detecting BTVs RNA in 310 blood samples. For real time and gel based RT-PCR segment-1 and segment-10 selected as conserve genes to search any BTV strains. Using these methods, 58 (%18.7) and 14 (%4.5) positive samples were detected among the clinically suspected sheep. Sensitivity of both molecular techniques evalua...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Journal of clinical pathology

دوره 56 10  شماره 

صفحات  -

تاریخ انتشار 2003