22 maggio 2022

Il famigerato laboratorio di Wuhan ha recentemente assemblato ceppi di vaiolo delle scimmie utilizzando metodi segnalati per la creazione di "agenti patogeni contagiosi"

L'Istituto di virologia di Wuhan ha assemblato un genoma del virus del vaiolo delle scimmie, consentendo l'identificazione del virus tramite test PCR, utilizzando un metodo che i ricercatori avevano segnalato come potenzialmente in grado di creare un "agente patogeno contagioso", rivela The National Pulse.

Lo studio è stato pubblicato per la prima volta nel febbraio 2022, pochi mesi prima dell'ultima epidemia internazionale di casi di vaiolo delle scimmie che sembra aver raggiunto gli Stati Uniti.

L'articolo, scritto da nove ricercatori dell'Istituto di virologia di Wuhan e pubblicato sulla rivista scientifica trimestrale Virologica Sinica del laboratorio, segue anche l'uso su larga scala dei test di reazione a catena della polimerasi (PCR) per identificare i soggetti positivi al COVID-19.

Nel documento i ricercatori sembrano aver identificato una porzione del genoma del virus del vaiolo delle scimmie, consentendo ai test PCR di identificare il virus: “Efficient Assembly of a Large Fragment of Monkeypox Virus Genome as a qPCR Template Using Dual-Selection Based Transformation-Associated Recombination.”("Assemblaggio efficiente di un ampio frammento del genoma del virus del vaiolo delle scimmie come templato qPCR utilizzando la ricombinazione associata alla trasformazione basata sulla doppia selezione").

I virus del vaiolo delle scimmie - indicati come "MPXV" nel documento - hanno ceppi "più patogeni e [sono] stati segnalati per infettare gli esseri umani in varie parti del mondo".

"La reazione a catena della polimerasi quantitativa (qPCR) è il gold standard per il rilevamento degli orthopoxvirus (compresi gli MPXV). Per il rilevamento dei pan-ortovirus, il gene E9L (DNA polimerasi) ha dimostrato di essere un eccellente bersaglio per i saggi qPCR. Per il rilevamento dell'MPXV, Li et al. hanno riferito che il gene C3L (proteina legante il complemento) potrebbe essere usato come bersaglio della qPCR per il ceppo MPXV del bacino del Congo", spiega il documento, prima di notare che la Cina non dispone di informazioni genetiche sufficienti sul virus per il rilevamento mediante PCR:

"Poiché l'infezione da MPXV non è mai stata associata a un focolaio in Cina, il materiale genomico virale necessario per la rilevazione mediante qPCR non è disponibile. In questa relazione, abbiamo impiegato la TAR bi-selettiva per assemblare un frammento genomico di MPXV di 55 kb che comprende E9L e C3L, due preziosi target per la qPCR per rilevare MPXV o altri orthopoxvirus".

"Lo scopo principale dell'assemblaggio di un frammento del genoma dell'MPXV è fornire un template nucleotidico per il rilevamento dell'MPXV", ha ribadito lo studio, che si è basato sul processo di ricombinazione associata alla trasformazione (TAR) per isolare un frammento genomico del virus del vaiolo delle scimmie.

"Come strumento efficiente per l'assemblaggio di grandi frammenti di DNA fino a 592 kb di lunghezza, l'assemblaggio TAR è diventato essenziale per la preparazione di cloni infettivi di grandi virus DNA/RNA", hanno spiegato i ricercatori.

Il documento riconosce che il TAR "applicato alla ricerca virologica potrebbe anche sollevare potenziali problemi di sicurezza, soprattutto quando il prodotto assemblato contiene un set completo di materiale genetico che può essere recuperato in un agente patogeno contagioso".

"In questo studio, sebbene un genoma virale completo sarebbe il modello di riferimento ideale per rilevare l'MPXV mediante qPCR, abbiamo cercato di assemblare solo un frammento virale di 55 kb, meno di un terzo del genoma dell'MPXV. Questo prodotto di assemblaggio è a prova di errore, eliminando virtualmente qualsiasi rischio di recupero in un virus infettivo e fornendo allo stesso tempo molteplici bersagli qPCR per rilevare l'MPXV o altri orthopoxvirus", hanno affermato i ricercatori.

Lo studio scoperto segue l'Istituto di virologia di Wuhan che ha condotto ricerche simili su ceppi di coronavirus dei pipistrelli in grado di infettare gli esseri umani, pur ammettendo che le sue strutture mancavano di adeguati protocolli di sicurezza di laboratorio.

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