Rapid detection of four non-fermenting Gram-negative bacteria directly from cystic fibrosis patient's respiratory samples on the BD MAX™ system

Talita T Rocchetti; Suzane Silbert; Alicia Gostnell; Carly Kubasek; Robert Jerris; Jennifer Vong; Raymond Widen

doi:10.1016/j.plabm.2018.e00102

Rapid detection of four non-fermenting Gram-negative bacteria directly from cystic fibrosis patient's respiratory samples on the BD MAX™ system

Pract Lab Med. 2018 May 24:12:e00102. doi: 10.1016/j.plabm.2018.e00102. eCollection 2018 Nov.

Authors

Talita T Rocchetti^{1

2}, Suzane Silbert¹, Alicia Gostnell¹, Carly Kubasek¹, Robert Jerris³, Jennifer Vong³, Raymond Widen¹

Affiliations

¹ Esoteric Testing Laboratory, Pathology Department, Tampa General Hospital, Tampa, FL, USA.
² Laboratório Especial de Microbiologia Clínica (LEMC), Federal University of São Paulo/UNIFESP, São Paulo, SP, Brazil.
³ Children's Healthcare of Atlanta, Atlanta, GA, USA.

Abstract

The aim of this study was to develop a multiplex PCR test to detect Achromobacter xylosoxidans (AX), Burkholderia cepacia (BC), Pseudomonas aeruginosa (PSA) and Stenotrophomonas maltophilia (SM) directly from CF patient's respiratory samples using the open mode of the BD MAX™ System. A total of 402 CF respiratory samples were evaluated by culture and PCR. Specific sets of primers and probes for each target were designed in-house. Out of 402 samples tested, 196 were identified as negative and 206 as positive by culture for AX, PSA, BC and SM. Among culture positive samples, PCR detected 21/27 AX, 4/5 BC, 138/140 PSA and 29/34 SM. In addition, PCR assay identified 35 samples as positive that were initially negative by culture for those 4 targets. The CF BDM test proved to be an excellent tool to detect AX, BC, PSA and SM by real-time PCR on an automated platform.

Keywords: Achromobacter xylosoxidana; BD Max; Burkholderia cepacia; Cystic Fibrosis; Pseudomonas aeruginosa; Stenotrophomonas maltophilia.