[{
  "type": "article-journal",
  "title": "LIPOPOLYSACHARIDE GENE BASED DEVELOPMENT AND OPTIMIZATION OF DIAGNOSTICS FOR FRANCISELLA TULARENSIS",
  "author": [
    {
      "family": "Muhammad",
      "given": ""
    },
    {
      "family": "Rabbani",
      "given": ""
    },
    {
      "family": "Muhammad",
      "given": ""
    },
    {
      "family": "Wasim",
      "given": ""
    },
    {
      "family": "Dewan",
      "given": ""
    },
    {
      "family": "Anjum",
      "given": ""
    },
    {
      "family": "Sheikh",
      "given": ""
    },
    {
      "family": "Ahmad",
      "given": ""
    },
    {
      "family": "Akhtar",
      "given": ""
    },
    {
      "family": "Nisar",
      "given": ""
    },
    {
      "family": "Jayarao",
      "given": ""
    },
    {
      "family": "Kirimanjeswara6.",
      "given": ""
    }
  ],
  "issued": {
    "date-parts": [[2017]]
  },
  "container-title": "Journal of Animal and Plant Sciences",
  "ISSN": "1018-7081",
  "volume": "27",
  "issue": "4",
  "page": "1161-1166",
  "DOI": "NA",
  "abstract": "<p><em>Francisella tularensis</em>&nbsp;(FT) is a Tier 1 select agent due to its low infectious dose and potential threat of being a bio-weapon while causing zoonotic disease in more than 150 mammalian species. Since there are concerns in its culturing at bio-containment deficient labs in developing countries and its subsequent diagnostics and surveillance accordingly, here we proposed a conventional PCR based test of choice that have been developed and optimized for detection of FT using&nbsp;<em>tul</em>4 gene specific primers. Annealing temperature and genomic DNA concentration was optimized using a live vaccine strain. Specificity of primer showed FT detection in soil DNA in the presence of other cross-reactive organism. Sensitivity was determined in two fold dilutions with detection limit of up to 320 pg/&micro;L. Utilizing pET28a vector, a construct was prepared containing transformed&nbsp;<em>tul</em>4 gene (450bp) showing 100% sequence homology to query gene sequence. These results suggest&nbsp;<em>tul</em>4 gene based constructs in vector could be shared to developing countries while developing diagnostic assays as positive control and could potentially be used for lipoprotein purification for ELISA kit development.</p>",
  "publisher": "Pakistan Agricultural Scientists Forum",
  "URL": "https://thejaps.org.pk/AbstractView.aspx?mid=2017-JAPS-146"
}]