Polymerase chain displacement reaction.

نویسندگان

  • Claire L Harris
  • Irma J Sanchez-Vargas
  • Ken E Olson
  • Luke Alphey
  • Guoliang Fu
چکیده

Quantitative PCR assays are now the standard method for viral diagnostics. These assays must be specific, as well as sensitive, to detect the potentially low starting copy number of viral genomic material. We describe a new technique, polymerase chain displacement reaction (PCDR), which uses multiple nested primers in a rapid, capped, one-tube reaction that increases the sensitivity of normal quantitative PCR (qPCR) assays. Sensitivity was increased by approximately 10-fold in a proof-of-principle test on dengue virus sequence. In PCDR, when extension occurs from the outer primer, it displaces the extension strand produced from the inner primer by utilizing a polymerase that has strand displacement activity. This allows a greater than 2-fold increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR. Increased sensitivity in PCDR would be useful in nucleic acid detection for viral diagnostics.

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

Comparison of fine needle aspiration and direct skin smear in the diagnosis of acute cutaneous leishmaniasis: polymerase chain reaction as a reference method

Background: Accurate diagnosis of cutaneous leishmaniasis wouldavoid unnecessary treatment and scar formation. Direct smear isthe most common method for the diagnosis of this disease but itsnegative result could not rule out the infection; so, the need formore sensitive methods is obvious. We conducted this study tocompare the efficiency of direct skin smear with smears preparedby fine needle a...

متن کامل

Polymerase Chain Reaction Study of Anogenital Condylomas for High Risk HPV Types

Background: Anogenital wart is a common sexually transmitted disease affecting both genders. The aim of this study was to evaluate human papilloma virus types of anogenital warts with Polymerase Chain Reaction (PCR). Method: 65 formalin fixed paraffin embedded (FFPE) tissue blocks of the anogenital warts obtained from the pathology lab of Loghman Hakim hospital were included in this study. Resu...

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:
  • BioTechniques

دوره 54 2  شماره 

صفحات  -

تاریخ انتشار 2013