Infectious pathogen detection arrays: viral detection in cell lines and postmortem brain tissue.

نویسندگان

  • Concepcion Conejero-Goldberg
  • Ena Wang
  • Chuli Yi
  • Terry E Goldberg
  • Lorraine Jones-Brando
  • Francesco M Marincola
  • Maree J Webster
  • E Fuller Torrey
چکیده

A unique array-based pathogen chip has been developed for the detection of viral RNA or DNA relevant to pathologies of the central nervous system. A total of 715 unique oligonucleotides (60-mer) representing approximately 100 pathogens were designed based on open reading frames (ORFs) from highly conserved and heterogenic regions within viral families. In addition, viral genes reflecting different stages of pathogen infection were also included to potentially define the stage of the viral infection. Viruses (double-stranded DNA, double- or single-stranded RNA, delta, retroid), parasites, and bacteria were included. Test samples labeled with Cy5 were examined by cohybridization with a reference RNA, labeled with Cy3, to the pathogen microarray chip. Good reproducibility of experiments was observed, based on data generated from duplicate hybridizations and duplicate spots on the microarray platform. A viral transcript detection sensitivity of 1 x 10(3) plaque-forming units (pfus) was achieved using selected cell lines and viruses. These findings suggest that the array-based platform described here is capable of detecting a broad spectrum of viruses in a single assay with relatively high sensitivity, specificity, and reproducibility. This method may be used to provide evidence of viral infection in postmortem tissue from psychiatric patients as well as a wide range of other diagnostic categories.

برای دانلود رایگان 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

Using Real-Time PCR to Detect Notifiable Viral Diseases (IPN, VHS and IHN) in Specific Pathogen Free (SPF) Rainbow Trout Broodstock Generations

The aim of this study was to identify different virulence pathogens using Real-Time PCR method for rapid and accurate detection of common viral diseases including Infectious Hematopoietic Necrosis (IHN), Viral Hemorrhagic Septicemia (VHS) and Infectious Pancreatic Necrosis disease (IPN) in rainbow trout broodstock have been the main generation of reproductive populations maintained in the natio...

متن کامل

Chicken infectious anaemia virus infection among broiler chicken flocks in Iran

Chicken infectious anaemia is a viral disease in young chickens which characterized by aplastic anaemia and immunosuppression. Between January 2004 and July 2005, an unusual hemorrhage in subcutaneous and intramuscular tissues of broiler chickens at slaughter houses of Mashhad, Isfahan and Tehran provinces were occurred. Postmortem examination revealed severe hemorrhages in the wings and muscle...

متن کامل

Development of a recombinant protein-based dot-blot hybridization assay for the detection of antibody to chicken infectious bronchitis virus

Nucleocapsid (N) protein of infectious bronchitis virus (IBV), one of the viral structural proteins, inducesstrong antibody response in natural infection. In this study, a simple, recombinant N protein-based dot-blottest was developed to serologically examine chicken serum samples for the presence of IBV antibody.Initially, 72 serum samples were tested for the presence of IBV antibody using a c...

متن کامل

Characterization of the BHK-21C5 Cell line and Its Introduction for use in Research, Diagnostics and Production of Biological Products

Background and Objectives: Several studies have been carried out on the use of cell lines in researches, production and processing of drugs and biological products, and on the identification of toxicity and efficacy.The present study was conducted to determine the characteristics of BHK-21 cell lines as a suitable substrate for use in vaccine production and quality control, viral culture and re...

متن کامل

Differentiation of Virulent and Non-Virulent Newcastle Disease Virus Isolates Using RT-PCR

Newcastle disease is one of the main concerns of poultry farmers. Detection of virulent strains of Newcastle disease virus (NDV) has a great impact on control measures against the disease. In this study RT-PCR was optimized in high sensitivity in order to differentiate the virulent from non-virulent NDV isolates directly in tissue homogenates. The vaccinal NDV strain and known field isolates we...

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

عنوان ژورنال:
  • BioTechniques

دوره 39 5  شماره 

صفحات  -

تاریخ انتشار 2005