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Journal of Clinical Microbiology, June 2005, p. 2616-2623, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2616-2623.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development, Technical Performance, and Clinical Evaluation of a NucliSens Basic Kit Application for Detection of Enterovirus RNA in Cerebrospinal Fluid

Christine C. Ginocchio,^1,2* Frank Zhang,¹ Amisha Malhotra,^3, Ryhana Manji,¹ Peter Sillekens,⁴ Helma Foolen,⁴ Marlieke Overdyk,⁴ and Margot Peeters⁴

North Shore-Long Island Jewish Health System Laboratories, Department of Molecular Diagnostics, Lake Success, New York,¹ North Shore University Hospital, Department of Laboratory Medicine,² North Shore University Hospital, Department of Pediatrics, Manhasset, New York,³ bioMérieux, Boxtel, The Netherlands⁴

Received 13 November 2004/ Returned for modification 19 January 2004/ Accepted 4 March 2005

The combination of nucleic acid sequence-based amplification and electrochemiluminescence detection was used to develop an internally controlled, highly sensitive and specific assay for the detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF). The analytical performance of the assay was determined using both in vitro-transcribed EV RNAs and viral culture isolates. The sensitivity of the assay was 10 EV RNA copies per amplification reaction. The assay detected all enteroviral isolates tested with no cross-reactivity to 21 nonenteroviral species, including rhinovirus and parechovirus. The clinical performance of the assay was evaluated by testing 992 CSF specimens collected from adult and pediatric patients. NucliSens EV results from a subset of 327 CSF samples were compared to viral culture of nasopharyngeal specimens and rectal swabs (n = 195) and/or CSF (n = 212). Of the 212 CSF samples, 96 samples were positive by either the NucliSens EV assay (94/96; 97.9%) or culture (63/96; 65.6%), and 61/96 (63.5%) were positive by both methods. The inclusion of an EV-specific internal control monitored the entire process, including the efficiency of nucleic acid extraction, amplification, and detection. In total, only five blood-clotted CSF samples (0.5%) were inhibited. The NucliSens EV assay demonstrated superior sensitivity over viral culture (P < 0.001), excellent specificity, clear delineation of positive samples, and minimal amplification inhibition.

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* Corresponding author. Mailing address: North Shore Long Island Jewish Health System Laboratories, 10 Nevada Drive, Lake Success, NY 11042. Phone: (516) 719-1079. Fax: (516) 719-1254. E-mail: cginocch{at}nshs.edu.

Present address: University of Medicine and Dentistry of New Jersey, Robert Woods Johnson Medical School, New Brunswick, NJ 08903.

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Journal of Clinical Microbiology, June 2005, p. 2616-2623, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2616-2623.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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