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<jats:p><jats:named-content content-type="genus-species">Streptococcus pneumoniae</jats:named-content>is both a commensal and a major pathogen that causes invasive disease in people of all ages. The introduction of serotype-specific pneumococcal vaccines has reduced the burden of disease but has also led to replacement with new strains; thus, serotyping remains important for vaccine-related disease surveillance. Conventional serotyping methods are laborious and expensive. We developed an easy-to-perform genotypic TaqMan array card (TAC) to identify<jats:named-content content-type="genus-species">S. pneumoniae</jats:named-content>strains, including<jats:italic>lytA</jats:italic>-based sequences, and 53 sequence-specific PCRs to identify 74 serotypes/serogroups covering all current vaccine types as well as prevalent nonvaccine types. The TAC method was evaluated on 146 clinical<jats:named-content content-type="genus-species">S. pneumoniae</jats:named-content>isolates and 13 nonpneumococcal species that naturally inhabit the upper respiratory tract and yielded 97% (142/146) sensitivity and 100% (13/13) specificity versus results of standard Quellung serotyping. The calculated limit of detection was 20 to 200 fg (∼8 to 84 genome equivalents) per reaction. On 23 blinded nasopharyngeal specimens that were pneumococcus culture positive, the TAC pan-pneumococcus<jats:italic>lytA</jats:italic>assay was positive in 21 (91% sensitivity versus culture). On TAC<jats:italic>lytA</jats:italic>-positive specimens, a serotype result was obtained on 86%, and the result was 95% accurate versus the subsequent culture's Quellung result. TAC also detected mixed serotypes in two specimens where Quellung detected only the predominant serotype. This TAC method yields fast and comprehensive serotyping compared to the standard method and may be useful on direct specimens.</jats:p>

Original publication

DOI

10.1128/jcm.00613-16

Type

Journal

Journal of Clinical Microbiology

Publisher

American Society for Microbiology

Publication Date

07/2016

Volume

54

Pages

1842 - 1850