Thermophilic helicase-dependent DNA amplification using the IsoAmp™ SE experimental kit for rapid detection of Streptococcus equi subspecies equi in clinical samples.

نویسندگان

  • Sergey Artiushin
  • Yanhong Tong
  • John Timoney
  • Bertrand Lemieux
  • Anne Schlegel
  • Huimin Kong
چکیده

A simple and portable assay for detection of Streptococcus equi subspecies equi has been developed based on amplification of S. equi-specific sequence using a thermophilic helicase-dependent reaction followed by visual detection of the amplicon in a disposable lateral flow cassette. An experimental kit (IsoAmp™ SE) was evaluated. Analytical sensitivity was 50 copies of S. equi genomic DNA per reaction. The IsoAmp SE assay had 100% specificity when applied to nasal swabs and washes. The assay was more sensitive than culture but less sensitive than nested polymerase chain reaction (PCR). The test requires neither expensive equipment nor extensive training of personnel, provides a practical alternative to culture or PCR assays for detection of S. equi in clinical samples, and expedites identification of atypical colonies of S. equi and Streptococcus zooepidemicus in the laboratory.

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

Development of a Loop-Mediated Isothermal Amplification Method for Detecting Streptococcus equi subsp. zooepidemicus and Analysis of Its Use with Three Simple Methods of Extracting DNA from Equine Respiratory Tract Specimens

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a dominant pathogenic bacterium in equine pneumonia. We developed a specific loop-mediated isothermal amplification (LAMP) method, which targets the gene encoding sorbitol-6-phosphate 2-dehydrogenase (sorD), for detecting S. zooepidemicus and examined the clinical efficacies of its use in combination with each of 3 DNA extraction met...

متن کامل

Development and application of loop-mediated isothermal amplification methods targeting the seM gene for detection of Streptococcus equi subsp. equi.

Loop-mediated isothermal amplification (LAMP) constitutes a potentially valuable diagnostic tool for rapid diagnosis of contagious diseases. In this study, we developed a novel LAMP method (seM-LAMP) to detect the seM gene of Streptococcus equi subsp. equi (S. equi), the causative agent of strangles in equids. The seM-LAMP successfully amplified the target sequence of the seM gene at 63°C withi...

متن کامل

Molecular and microscopic detection of Theileria equi and Babesia caballi in horses in Kurdestan Province, Iran

Equine piroplasmosis is a tick-borne disease caused by intra-erythrocyte protozoa, Theileria equi and Babesia caballi. The present study aimed to detect piroplasm infection in horses in Kurdestan Province, Iran, through molecular and microscopic approaches.n this study, 186 blood samples were randomly collected from horses of five regions of Kurdestan Province. The Tbs-S/Tbs-A primer set was us...

متن کامل

Molecular detection of equine piroplasms in donkeys (Equus asinus) in North Khorasan province, Iran

Equine piroplasmosis is a tickborne disease of equids with worldwide distribution, caused by Theileria equi and Babesia caballi. The aim of this study was molecular detection of T. equi and B. caballi in donkeys in northeastern Iran and investigate the association between positivity of piroplasm infection and host-related factors. In the present study, Blood samples were collected from 106 appa...

متن کامل

Detection of Streptococcus equi subspecies equi using a triplex qPCR assay

Genome sequencing data for Streptococcus equi subspecies equi and zooepidemicus were used to develop a novel diagnostic triplex quantitative PCR (qPCR) assay targeting two genes specific to S. equi (eqbE and SEQ2190) and a unique 100 base pair control DNA sequence (SZIC) inserted into the SZO07770 pseudogene of S. zooepidemicus strain H70. This triplex strangles qPCR assay can provide results w...

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 veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

دوره 23 5  شماره 

صفحات  -

تاریخ انتشار 2011