Analytical and clinical validation of a novel, laboratory-developed, modular multiplex-PCR panel for fully automated high-throughput detection of 16 respiratory viruses.
J Clin Virol
; 173: 105693, 2024 08.
Article
em En
| MEDLINE
| ID: mdl-38820916
ABSTRACT
BACKGROUND:
Viral respiratory Infections pose a health risk, especially to vulnerable patient populations. Effective testing programs can detect and differentiate these infections at an early stage, which is particularly important for high-risk clinical departments. The objective of this study was to develop and validate a multiplex PCR-panel for 16 different respiratory viruses on a fully-automated high-throughput platform.METHODS:
Three multiplex-PCR assays were designed to run on the cobas5800/6800/8800 systems, consolidating 16 viral targets RESP1 SARS-CoV-2, influenza-A/B, RSV; RESP2 hMPV, hBoV, hAdV, rhino-/ENV; RESP3 HPIV-1-4, hCoV-229E, hCoV-NL63, hCoV-OC43, hCoV-HKU1. Analytic performance was evaluated using digital-PCR based standards and international reference material. Clinical performance was determined by comparing results from clinical samples with reference assays.RESULTS:
Analytical sensitivity (i.e. lower limit of detection (LoD), 95 % probability of detection) was determined as follows SARS-CoV-2 29.3 IU/ml, influenza-A 179.9 cp/ml, influenza-B 333.9 cp/ml and RSV 283.1 cp/ml. LoDs of other pathogens ranged between 9.4 cp/ml (hCoV-NL63) and 21,419 cp/ml (HPIV-2). Linearity was verified over 4-7 log-steps with pooled standard differentials (SD) ranging between 0.18-0.70ct. Inter-/intra-run variability (precision) was assessed for all targets over 3 days. SDs ranged between 0.13-0.74ct. Positive agreement in clinical samples was 99.4 % and 95 % for SARS-CoV-2 and influenza-A respectively. Other targets were in the 80-100 % range. Negative agreement varied between 96.3-100 %.DISCUSSION:
Lab-developed tests are a key factor for effective clinical diagnostics. The multiplex panel presented in this study demonstrated high performance and provides an easily scalable high-throughput solution for respiratory virus testing, e.g. for testing in high-risk patient populations.Palavras-chave
Texto completo:
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Infecções Respiratórias
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Sensibilidade e Especificidade
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Reação em Cadeia da Polimerase Multiplex
Limite:
Humans
Idioma:
En
Revista:
J Clin Virol
Ano de publicação:
2024
Tipo de documento:
Article