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PathogenMip assay: a multiplex pathogen detection assay

Michael S Akhras1, Sreedevi Thiyagarajan, Andrea C Villablanca

  • 1Stanford Genome Technology Center, Stanford University, Palo Alto, California, United States of America.

Plos One
|February 22, 2007
PubMed

Insights

The novel PathogenMip Assay offers a cost-effective method for pathogen detection using highly discriminating probes. This assay accurately genotypes human papillomavirus (HPV), showing potential for broad microbial identification.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • The Molecular Inversion Probe (MIP) assay is established for human SNP detection.
  • Pathogen detection requires precise and accessible genotyping methods.

Purpose of the Study:

  • To introduce the PathogenMip Assay, a comprehensive protocol for pathogen detection.
  • To develop a cost-effective method for producing highly discriminating probes for genotyping.

Main Methods:

  • Developed a novel, ligase-based protocol for probe production.
  • Targeted 24 human papillomavirus (HPV) genotypes associated with cervical cancer.
  • Validated the assay using pyrosequencing, microarray detection, and conventional sequencing.

Main Results:

  • Achieved 100% agreement with conventional sequencing for HPV genotyping in tumor DNA.
  • Demonstrated high sensitivity and selectivity using HPV plasmids.
  • Successfully genotyped clinically relevant HPV genotypes.

Conclusions:

  • The PathogenMip Assay provides a widely accessible protocol for pathogen genotyping.
  • The assay is experimentally validated and shows potential for detecting and characterizing diverse microbes.

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