Evaluation of Four Commercial Multiplex Molecular Tests for the Diagnosis of Acute Respiratory Infections

Abstract

Acute Respiratory Infections (ARIs) are responsible for considerable morbidity and mortality worldwide. Documentation of respiratory specimens can help for an appropriate clinical management with a significant effect on the disease progress in patient, the antimicrobial therapy used and the risk of secondary spread of infection. Here, we compared the performances of four commercial multiplex kits used in French University Hospital diagnostic microbiology laboratories for the detection of ARI pathogens (i.e., the xTAG Respiratory Viral Panel Fast, RespiFinder SMART 22, CLART PneumoVir and Fast Track Diagnostics Respiratory Pathogen 33 kits). We used a standardised nucleic acids extraction protocol and a comprehensive comparative approach that mixed reference to well established real-time PCR detection techniques and analysis of convergent positive results. We tested 166 respiratory clinical samples and identified a global high degree of correlation for at least three of the techniques (xTAG, RespiFinder and FTD33). For these techniques, the highest Youden's index (YI), positive predictive (PPV) and specificity (Sp) values were observed for Core tests (e.g., influenza A [YI:0.86-1.00; PPV:78.95-100.00; Sp:97.32-100.00] & B [YI:0.44-1.00; PPV:100.00; Sp:100.00], hRSV [YI:0.50-0.99; PPV:85.71-100.00; Sp:99.38-100.00], hMPV [YI:0.71-1.00; PPV:83.33-100.00; Sp:99.37-100.00], EV/hRV [YI:0.62-0.82; PPV:93.33-100.00; Sp:94.48-100.00], AdV [YI:1.00; PPV:100.00; Sp:100.00] and hBoV [YI:0.20-0.80; PPV:57.14-100.00; Sp:98.14-100.00]). The present study completed an overview of the multiplex techniques available for the diagnosis of acute respiratory infections.

Fig 1. Decision tree (algorithm used for Core tests).

(Inf A: Influenza A, Inf B: Influenza B, hRSV: human Respiratory Syncytial Virus, hMPV: human Metapneumovirus, EV + hRV:; Enterovirus + human Rhinovirus, AdV: Adenovirus and hBoV: human Bocavirus).

Fig 2. Estimate of detection limit of the different kits.

Inf A H1N1: Influenza A H1N1 (A/Marseille/9410380/2009), Inf A H3N2: Influenza A H3N2 (A/Marseille/4781598/2009), Inf B: Influenza B (B/Marseille/4461097/2008), Inf C: Influenza C (C/Leningrad:232/83), hRSV A and B: human Respiratory Syncytial virus type A (Long) and B (Gb2), hRV: Rhinovirus (MAR2007 3813047), EV Cox B3: Enterovirus (Human Coxsackievirus B3 clinical strain B32679), hPeV: human Parechovirus type 3 (UNK), AdV B: Adenovirus type B (MAR2007 7416184 clinical strain), and hMPV A: human Metapneumovirus type A.