Development of High-throughput Liquid Chips for Respiratory Virus Detection

A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses

“…Technically, Luminex xMAP analysis has higher sensitivity and specificity than real-time PCR since the liquid-phase hybridization after amplification can decrease the background interference and two-fluorescence detection can increase the specificity [28,36]. Through the study, we believe that we can make the rMLA assay better with some improvements.…”

“…No obvious difference was also found between the both, and in fact the great consistency of the diagnostic performance presented in the detection of clinical specimens(accordance rates > 95%, Kappa values > 0.9 for all targets). Some reports had the similar result [36,37], but there were also reports that Luminex xMAP analysis for some target was more or less sensitive than real-time PCR [20,37]. This mostly depended on the primer design and reaction optimization of the assay according to our experiences.…”

“…And the primer/ probe design for a Luminex assay becomes easier because the criterion of probe and the size of amplicon are not as strict as a real-time PCR. However, the product should be less than 300 bp to assure the amplification efficiency and analytical sensitivity [36]. In this study, the same primers and probes (slightly modified) shared by the rMLA and the real-time RT-PCR ensured not only the amplification efficiency and sensitivity of the rMLA assay, but also ensured the comparability of the two assays.…”

“…Its sensitivity and specificity can be improved by asymmetric multiplex PCR, adopting a protocol of differential cycle conditions based on temperature differential primer design [20,39,40]. The extra exonuclease I can be added to eliminate the residual primers before hybridization [11,36]. So, the overall turnaround time of the assay may increase a little (less than 30 minutes).…”

Bocavirus Humano 1: su participación en la infección respiratoria aguda y epidemiología en Argentina

“…Alternativamente, se contempla la ocurrencia de reinfecciones frecuentes durante la niñez, postulándose que sólo la primoinfección estaría ligada a manifestaciones clínicas (35,43) . A pesar los numerosos trabajos en niños (o tal vez precisamente porque todos los esfuerzos se concentraron inicialmente en dicha población), y debido a que aún es muy limitada la información sobre la patogénesis asociada a la infección, el HBoV1 no se investiga de rutina en el contexto de la práctica clínica -sí en centros médicos de países desarrollados, donde incluso existen ensayos diagnósticos disponibles comercialmente (45). Sin embargo, en los últimos años se ha ampliado el campo de estudio y recientemente se han reportado casos graves con desorden neurológico y fatales, tanto en niños como en adultos (46)(47)(48)(49) .…”