<div dir="ltr"><br clear="all"><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">Dear colleagues,</font></span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">We are pleased to announce that the publication
of our article is now available online:</font></span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 0pt"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">Qingzhong
Wu, <span style="color:black">Katherine</span> C. Prager, Tracey Goldstein, David
P. Alt, Renee L. Galloway, Richard L. Zuerner, James O. Lloyd-Smith and Lori
Schwacke. 2014.<sup> </sup>Development of a real-time PCR for the detection of
pathogenic <span> </span><i>Leptospira</i> spp. in <span style="color:black">California sea lions. Diseases
of Aquatic Organisms. </span>DOI: <span style="color:black">10.3354/dao02752</span></font></span><span style="color:black;font-family:"Helvetica","sans-serif""></span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt"><a href="http://www.int-res.com/abstracts/dao/v110/n3/p165-172/"><font color="#0000ff">http://www.int-res.com/abstracts/dao/v110/n3/p165-172/</font></a></span></p>
<font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt"> </span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt">Abstract:</span><span style="font-family:"Times New Roman","serif";font-size:12pt"></span></p>
<font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal;text-indent:0.5in" class="MsoNormal"><font color="#000000"><span style="font-family:"Times New Roman","serif";font-size:12pt">Several real-time PCR assays are currently used for detection of
pathogenic <i>Leptospira</i> spp.; however, </span><span style="font-family:"Times New Roman","serif";font-size:12pt">few methods have been described for the successful evaluation of
clinical urine samples.<span>  </span></span><span style="font-family:"Times New Roman","serif";font-size:12pt">This study reports
a rapid assay for the detection of pathogenic <i>Leptospira </i>spp.<i> </i>in <span style="color:black">California sea lions </span>(<i>Zalophus californianus</i><span style="color:black">) </span>using real-time
PCR with primers and a probe targeting the <i><span style="color:black">lipL32</span></i> gene.<span> 
</span>The PCR assay had high analytic sensitivity - the limit of detection was
three genome copies per PCR volume using </span><i><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt">L.</span></i><i><span style="font-family:"Times New Roman","serif";font-size:12pt"> interrogans</span></i><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt"> serovar Pomona DNA</span><span style="font-family:"Times New Roman","serif";font-size:12pt">; and <span style="color:black">100% analytic
specificity - it</span> detected all pathogenic leptospiral serovars tested and
none of the non-pathogenic <i>Leptospira</i>
species <i>L. biflexa </i>and</span><i><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt">
L. meyeri</span></i><span style="color:black;font-family:"Times New Roman","serif";font-size:12pt"> serovar Semaranga</span><i><span style="font-family:"Times New Roman","serif";font-size:12pt">,</span></i><span style="font-family:"Times New Roman","serif";font-size:12pt"> the intermediate species <i>L. inadai</i>, or the non-<i>Leptospira</i> pathogens tested.<span>  </span>Our assay had an amplification efficiency of
1.00.<span>  </span>Comparisons between the real-time
PCR assay and culture isolation for detection of pathogenic <i>Leptospira </i>spp<i>. </i>in <span style="color:black">urine and kidney tissue samples from
California sea lions </span>showed that samples were more often positive by real-time
<span style="color:black">PCR than by culture methods.<span>  </span></span>Inclusion of an internal amplification
control in the real-time PCR assay showed no inhibitory effects in <span style="color:black">PCR negative</span> samples.<span>  </span>These <span style="color:black">studies indicated
that our real-time PCR assay has high analytic sensitivity and specificity for the
rapid detection of pathogenic </span><i>Leptospira
</i>species <span style="color:black">in </span>urine and kidney tissue samples.</span></font></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000"> </font></span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">Best regards,</font></span></p><font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">Qingzhong (David) Wu</font></span></p>
<font color="#000000" size="3" face="Times New Roman">

</font><p style="margin:0in 0in 10pt;line-height:normal" class="MsoNormal"><span style="font-family:"Times New Roman","serif";font-size:12pt"><font color="#000000">Molecular microbiologist</font></span></p>
<font color="#000000" size="3" face="Times New Roman">

</font><br>-- <br><pre cols="72"><font face="comic sans ms,sans-serif">Qingzhong (David) Wu, <font face="Trebuchet MS">JHT, Inc. - Contractor</font> 

Hollings Marine Laboratory
National Ocean Service
331 Ft. Johnson Rd.
Charleston, SC 29412

Office 843-762-8940
</font><a href="mailto:david.wu@noaa.gov" target="_blank"><font face="comic sans ms,sans-serif">david.wu@noaa.gov</font></a>
</pre>
</div>