A palm-sized monkeypox POCT nucleic acid detection technology!
Research Summary
Outbreaks of monkeypox (formerly known as Monkeypox) have been declared a public health emergency of international concern. However, traditional polymerase chain reaction (PCR) diagnostic techniques are not ideal for field application.
To perform sample-to-result Mpox viral particle testing outside the laboratory, the authors developed an easy-to-handle, palm-sized pouch called the Mpox Home Self-Test and Stuck Pouch (MASTR Pouch).
In this handheld laboratory (MASTR Pouch), rapid and accurate visual detection was achieved by combining recombinant polymerase amplification (RPA) with the CRISPR/Cas12a system. From viral particle lysis to visual readout, the MASTR Pouch requires only four simple steps to complete the analysis process within 35 minutes. 53 Mpox pseudovirion particles (10.6 particles/μL) could be detected in the exudate.
To test the utility, 104 simulated Mpox clinical exudate specimens were tested. Clinical sensitivity was determined to be 91.7%-95.8%. No false positive results occurred and 100% clinical specificity was verified. MASTR Pouch is close to the World Health Organization Point-of-Care Diagnostic Criteria (ASSURD), which will be beneficial in mitigating the global spread of Mpox. The versatility of the MASTR Pouch may further revolutionize infection diagnostics.
key Technology
CRISPR technology; infectious disease; comprehensive detection tool; Mpox; POCT diagnosis; recombinase polymerase amplification
a) Schematic diagram of the general principle of RPA-CRISPR/Cas12a comprehensive detection;
b) Simple operation of detecting Mpox virus using the palm-sized MASTR Pouch.
a) The main components in MASTR Pouch, including mini straw, ready-to-use tube, sampling tube, mini power box, automatic readout device (ARD) and MHP30 mini constant temperature heating block;
b) the internal structure of the ARD;
c) Detection of different copies of Mpox B6R plasmid in normal saline (from 6. 9×104 to 0 copies);
d) Representative images obtained by detecting different numbers of B6R-type Mpox pseudoviruses (ranging from 5.3 × 104 to 0 virus particles) in normal saline;
e) Representative images obtained from the detection of different numbers of B6R-type Mpox pseudoviruses (ranging from 5.3 × 104 to 0 virus particles) in the exudate matrix.
a, b) Confusion matrix comparing the results of standard qPCR assays and MASTR Pouch-based assays. The preparation and detection of simulated clinical specimens followed the triple-blind rule. The target gene was (a) B6R gene or (b) F3L gene.
c) Correlation between positive results of MASTR Pouch-based assays and Ct values of qPCR assays.
