tetano
Editor, Senior Moderator
J Clin Microbiol. 2011 Dec 28. [Epub ahead of print]
Pooling Nasopharyngeal/Throat Swab Specimens to Increase Testing Capacity for Influenza Viruses by PCR.
Van TT, Miller J, Warshauer DM, Reisdorf E, Jernigan D, Humes R, Shult PA.
Source
Wisconsin State Laboratory of Hygiene, University of Wisconsin - Madison, Madison, WI.
Abstract
Real-time PCR methodology can be applied to rapidly and accurately detect influenza viruses. During times of surge testing or enhanced pandemic surveillance, public health laboratories (PHLs) may experience overwhelming demand for testing, even while the prevalence of positive specimens remains low. To improve surge laboratory capacity and testing efficiency, we evaluated whether nasopharyngeal (NP)/throat swab specimens can be pooled and tested for the presence of the 2009 H1N1 influenza virus without a reduction in sensitivity. Pools of ten specimens were extracted and concentrated upon elution on the MagNA Pure LC instrument, and real-time PCR was performed on the Applied Biosystems 7500 Fast platform, using the CDC Swine Influenza Virus Real-Time RT-PCR Detection Panel (rRT PCR Swine Flu Panel). Specimens in positive pools were singly re-extracted and re-tested by PCR to identify individual positive samples. Initial studies showed spiking a pool of nine negative specimens (100 μl each) or 900 μl of VTM with 100 μl of a positive clinical specimen caused no loss of sensitivity by rRT-PCR testing. Pools containing either multiple positive specimens or specimens positive for other respiratory viruses also showed no negative effect on C(t) values. To test the robustness of the pooling protocol, a panel of 50-blinded samples was sent to three PHLs and tested in five pools of ten. All PHLs correctly identified the positive specimens. This study demonstrates the feasibility of using a pooling strategy to increase capacity and conserve resources during surge testing, periods of enhanced influenza surveillance when the prevalence is low.
PMID:
22205820
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22205820
Pooling Nasopharyngeal/Throat Swab Specimens to Increase Testing Capacity for Influenza Viruses by PCR.
Van TT, Miller J, Warshauer DM, Reisdorf E, Jernigan D, Humes R, Shult PA.
Source
Wisconsin State Laboratory of Hygiene, University of Wisconsin - Madison, Madison, WI.
Abstract
Real-time PCR methodology can be applied to rapidly and accurately detect influenza viruses. During times of surge testing or enhanced pandemic surveillance, public health laboratories (PHLs) may experience overwhelming demand for testing, even while the prevalence of positive specimens remains low. To improve surge laboratory capacity and testing efficiency, we evaluated whether nasopharyngeal (NP)/throat swab specimens can be pooled and tested for the presence of the 2009 H1N1 influenza virus without a reduction in sensitivity. Pools of ten specimens were extracted and concentrated upon elution on the MagNA Pure LC instrument, and real-time PCR was performed on the Applied Biosystems 7500 Fast platform, using the CDC Swine Influenza Virus Real-Time RT-PCR Detection Panel (rRT PCR Swine Flu Panel). Specimens in positive pools were singly re-extracted and re-tested by PCR to identify individual positive samples. Initial studies showed spiking a pool of nine negative specimens (100 μl each) or 900 μl of VTM with 100 μl of a positive clinical specimen caused no loss of sensitivity by rRT-PCR testing. Pools containing either multiple positive specimens or specimens positive for other respiratory viruses also showed no negative effect on C(t) values. To test the robustness of the pooling protocol, a panel of 50-blinded samples was sent to three PHLs and tested in five pools of ten. All PHLs correctly identified the positive specimens. This study demonstrates the feasibility of using a pooling strategy to increase capacity and conserve resources during surge testing, periods of enhanced influenza surveillance when the prevalence is low.
PMID:
22205820
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22205820