Real-time PCR positivity for detection of Nm, Spn, and Hi in clinical samples received as part of Sao Paulo’s national surveillance for routine diagnosis analysis.

<p>Real-time PCR positivity for detection of Nm, Spn, and Hi in clinical samples received as part of Sao Paulo’s national surveillance for routine diagnosis analysis.</p

Values of sensitivity, specificity and Kappa index of real-time PCR assay using purified DNA from CSF impregnated on FTA^TM card.

<p>Values of sensitivity, specificity and Kappa index of real-time PCR assay using purified DNA from CSF impregnated on FTA^TM card.</p

Use of cerebrospinal fluid and serum samples impregnated on FTA^TM Elute filter paper for the diagnosis of infections caused by <i>Neisseria meningitidis</i>, <i>Streptococcus pneumoniae</i> and <i>Haemophilus influenzae</i>

<div><p>Background</p><p>The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTA^TM Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo’s national surveillance for routine diagnosis were selected for this study.</p><p>Methods</p><p>The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory’s standard DNA extraction, and processing the same samples after being dried and stored on FTA^TM card, and DNA extracted following the manufacturer’s instructions.</p><p>Results</p><p>The sensitivities for detection of <i>Neisseria meningitidis</i>, <i>Streptococcus pneumoniae</i>, and <i>Haemophilus influenzae</i> from CSF dried and stored on FTA^TM cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory’s standard methodology, results showed high concordance, with Kappa index ranges of 0.9877–1.00 for CSF, and 0.8004–1.00 for serum samples.</p><p>Conclusion</p><p>The use of FTA^TM cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost.</p></div

Processing algorithm of samples.

<p>Processing algorithm of samples.</p