<AbstractText>In September, 2017, human monkeypox re-emerged in Nigeria, 39 years after the last reported case. We aimed to describe the clinical and epidemiological features of the 2017-18 human monkeypox outbreak in Nigeria. We reviewed the epidemiological and clinical characteristics of cases of human monkeypox that occurred between Sept 22, 2017, and Sept 16, 2018. Data were collected with a standardised case investigation form, with a case definition of human monkeypox that was based on previously established guidelines. Diagnosis was confirmed by viral identification with real-time PCR and by detection of positive anti-orthopoxvirus IgM antibodies. Whole-genome sequencing was done for seven cases.
Haplotype analysis results, genetic distance data, and epidemiological data were used to infer a likely series of events for potential human-to-human transmission of the west African clade of monkeypox virus 122 confirmed or probable cases of human monkeypox were recorded in 17 states, including seven deaths (case fatality rate 6%). People infected with monkeypox virus were aged between 2 days and 50 years (median 29 years [IQR 14]), and 84 (69%) were male. All 122 patients had vesiculopustular rash, and fever, pruritus, headache, and lymphadenopathy were also common.
The rash affected all parts of the body, with the face being most affected. The distribution of cases and contacts suggested both primary zoonotic and secondary human-to-human transmission. Two cases of health-care-associated infection were recorded. Genomic analysis suggested multiple introductions of the virus and a single introduction along with human-to-human transmission in a prison facility.
This study describes the largest documented human outbreak of the west African clade of the monkeypox virus. Our results suggest endemicity of monkeypox virus in Nigeria, with some evidence of human-to-human transmission. Further studies are necessary to explore animal reservoirs and risk factors for transmission of the virus in Nigeria.
Characterization of Monkeypox virus dissemination in the black-tailed prairie dog (Cynomys ludovicianus) through in vivo bioluminescent imaging
Monkeypox virus (MPXV) is a member of the genus Orthopoxvirus, endemic in Central and West Africa. This viral zoonosis was introduced into the United States in 2003 via African rodents imported for the pet trade and caused 37 human cases, all linked to exposure to MPXV-infected black-tailed prairie dogs (Cynomys ludovicianus). Prairie dogs have since become a useful model of MPXV disease, utilized for testing of potential medical countermeasures. In this study, we used recombinant MPXV containing the firefly luciferase gene (luc) and in vivo imaging technology to characterize MPXV pathogenesis in the black-tailed prairie dog in real time.
West African (WA) MPXV could be visualized using in vivo imaging in the nose, lymph nodes, intestines, heart, lung, kidneys, and liver as early as day 6 post infection (p.i.). By day 9 p.i., lesions became visible on the skin and in some cases in the spleen. After day 9 p.i., luminescent signal representing MPXV replication either increased, indicating a progression to what would be a fatal infection, or decreased as infection was resolved. Use of recombinant luc+ MPXV allowed for a greater understanding of how MPXV disseminates throughout the body in prairie dogs during the course of infection. This technology will be used to reduce the number of animals required in future pathogenesis studies as well as aid in determining the effectiveness of potential medical countermeasures.
Preliminary Screening and In Vitro Confirmation of Orthopoxvirus Antivirals
The lack of antiviral drugs for the treatment of orthopoxvirus disease represents an unmet medical need, particularly due to the threat of variola virus (the causative agent of smallpox) as an agent of biowarfare or bioterrorism (Henderson, 283:1279-1282, 1999). In addition to variola, monkeypox, cowpox, and vaccinia viruses are orthopoxviruses of concern to human health (Lewis-Jones, 17:81-89, 2004). Smallpox vaccination, using the closely related vaccinia virus, is no longer provided to the general public leading to a worldwide population increasingly susceptible not only to variola but to monkeypox, cowpox, and vaccinia viruses as well.
Orthopoxviruses share similar life cycles (Fenner et al., WHO, Geneva, 1988), and significant nucleotide and protein homology, and are immunologically cross-protective against other species within the genus, which was the basis of the highly successful vaccinia virus vaccine. These similarities also serve as the basis for screening for antivirals for dangerous pathogens such as variola and monkeypox virus using generally safer viruses such as cowpox and vaccinia.
Methods for preliminary screening and initial characterization of potential orthopoxvirus antivirals in vitro, using vaccinia virus as a relatively safe surrogate for more pathogenic orthopoxviruses, are described herein. They include candidate identification in a viral cytopathic effect (CPE) assay as well as evaluation of the antiviral activity in inhibition assays to determine mean effective (or inhibitory) concentrations (EC50 or IC50). These assays were utilized in the identification and early characterization of tecovirimat (ST-246) (Yang et al., 79:13,139-13,149, 2005). These initial steps in identifying and characterizing the antiviral activity should be followed up with additional in vitro studies including specificity testing (for other orthopoxviruses and against other viruses), single-cycle growth curves, time of addition assays, cytotoxicity testing, and identification of the drug target.
Analgesia during Monkeypox Virus Experimental Challenge Studies in Prairie Dogs (Cynomys ludovicianus)
Because human patients with monkeypox virus (MPXV) infection report painful symptoms, it is reasonable to assume that animals infected with MPXV experience some degree of pain. Understanding whether and how analgesics affect MPXV disease progression is crucial when planning in vivo challenge experiments. In the current study, we challenged prairie dogs with a low dose (4 ×10³ pfu) of MPXV and treated with meloxicam (NSAID) or buprenorphine (opioid); control animals did not receive analgesia or received analgesia without MPXV challenge. Subsets of animals from each group were serially euthanized during the course of the study.
Disease progression and viral kinetics were similar between groups, but MXPVinfected, meloxicam-treated animals showed increasing trends of morbidity and mortality compared with other groups.
Differences between no-analgesia MPXV-infected control animals and MPXV-infected animals treated with buprenorphine were minimal. The findings in the current study allow more informed decisions concerning the use of analgesics during experimental MPXV challenge studies, thereby improving animal welfare. In light of these findings, we have modified our pain scale for this animal model to include the use of buprenorphine for pain relief when warranted after MPXV challenge.
Recombinase polymerase amplification assay for rapid detection of Monkeypox virus.
In this study, a rapid method for the detection of Central and West Africa clades of Monkeypox virus (MPXV) using recombinase polymerase amplification (RPA) assay targeting the G2R gene was developed. MPXV, an Orthopoxvirus, is a zoonotic dsDNA virus, which is listed as a biothreat agent. RPA was operated at a single constant temperature of 42°C and produced results within 3 to 10 minutes. The MPXV-RPA-assay was highly sensitive with a limit of detection of 16 DNA molecules/μl.
Monkeypox virus |
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Oneq-VH747-OneqVH747100D | Bioingentech | Oneq-VH747-100D |
Monkeypox virus |
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Oneq-VH747-OneqVH747150D | Bioingentech | Oneq-VH747-150D |
Monkeypox virus |
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Oneq-VH747-OneqVH74750D | Bioingentech | Oneq-VH747-50D |
Monkeypox Virus, 20 tests/kit |
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U202028-20 | USTAR Biotechnologies (Hangzhou) | test |
MonkeyPox Virus (MPV) ELISA Kit |
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SL0026Mk | Sunlong | - |
MonkeyPox Virus (MPV) ELISA Kit |
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YX-SL0036Mk | Sunlong | - |
Recombinant Monkeypox Virus antigen |
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E41H094 | EnoGene | 100ug |
Recombinant Monkeypox Virus antigen |
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E41H095 | EnoGene | 100ug |
Monkeypox Virus Real Time PCR Kit |
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PDPS-AR064 | Creative Biogene | 1 unit |
Monkeypox Virus Real Time PCR Kit |
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ZD-0076-01 | Gentaur Genprice | 25 tests/kit |
Monkeypox Virus Real Time PCR Kit |
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ZD-0076-02 | Gentaur Genprice | 25 tests/kit |
Monkeypox Virus Real Time PCR Kit |
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YJC70115NW-25T | Jiangsu Bioperfectus Technologies | 25 tests/kit |
Monkeypox Virus (MPXV) E8L ELISA Kit |
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RK04609 | Abclonal | 96T |
Monkeypox virus (MPXV) E8L Rabbit mAb |
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A23087 | Abclonal | 500μL |
Recombinant Monkeypox virus A29 Protein |
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RP01543 | Abclonal | 100μg |
Recombinant Monkeypox virus L1R Protein |
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RP01544 | Abclonal | 500μg |
Recombinant Monkeypox virus A29 Protein |
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RP01546 | Abclonal | 50μg |
Recombinant Monkeypox virus D6L Protein |
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RP01606 | Abclonal | 50μg |
Monkeypox Virus E8L (MPXV_USA_2022_MA001), mFc Tag |
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E24PVA019 | EnoGene | 50 μg |
Recombinant Monkeypox virus A30L Protein |
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RP01593 | Abclonal | 20μg |
Recombinant Monkeypox virus B16R Protein |
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RP01594 | Abclonal | 50μg |
PCR detection kit for Monkeypox Virus |
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DA1430 | Daan Gene | 24 test/kit |
Monkeypox Virus Detection Kit (RT-PCR) |
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SYNT-RTPCR-24 | Nanjing Synthgene Medical Technology | 24 test/kit |
Dynamiker Monkeypox Virus Ag Rapid Test |
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DNK-2114-1 | Dynamiker Biotechnology | 20 tests |
Mouse Anti Monkeypox Virus E8L Monoclonal Clone SAA0285 |
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IMSAMPXVE8LSAA0285C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus E8L Monoclonal Clone SAA0285 |
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IMSAMPXVE8LSAA0285C1MG | Innovative research | each |
Mouse Anti Monkeypox Virus H3L Monoclonal Clone SAA0282 |
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IMSAMPXVH3LSAA0282C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus H3L Monoclonal Clone SAA0282 |
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IMSAMPXVH3LSAA0282C1MG | Innovative research | each |
Mouse Anti Monkeypox Virus M1R Monoclonal Clone SAA0283 |
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IMSAMPXVM1RSAA0283C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus M1R Monoclonal Clone SAA0283 |
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IMSAMPXVM1RSAA0283C1MG | Innovative research | each |
Mouse Anti Monkeypox Virus M1R Monoclonal Clone SAA0284 |
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IMSAMPXVM1RSAA0284C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus M1R Monoclonal Clone SAA0284 |
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IMSAMPXVM1RSAA0284C1MG | Innovative research | each |
Mouse Anti Monkeypox Virus A29L Monoclonal Clone SAA0286 |
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IMSAMPXVA29LSAA0286C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus A29L Monoclonal Clone SAA0286 |
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IMSAMPXVA29LSAA0286C1MG | Innovative research | each |
Mouse Anti Monkeypox Virus A35R Monoclonal Clone SAA0287 |
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IMSAMPXVA35RSAA0287C100UG | Innovative research | each |
Mouse Anti Monkeypox Virus A35R Monoclonal Clone SAA0287 |
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IMSAMPXVA35RSAA0287C1MG | Innovative research | each |
Monkeypox Virus L1R (strain Zaire-96-I-16), His Tag |
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E24PVA017 | EnoGene | 50 μg |
Monkeypox Virus I1L (strain Zaire-96-I-16), His Tag |
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E24PVA018 | EnoGene | 50 μg |
Monkeypox Virus B6R (strain Zaire-96-I-16), His Tag |
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E24PVA020 | EnoGene | 50 μg |
Monkeypox Virus B6R (strain Zaire-96-I-16), mFc Tag |
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E24PVA021 | EnoGene | 50 μg |
Monkeypox Virus L1R (strain Zaire-96-I-16), mFc Tag |
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E24PVA025 | EnoGene | 50 μg |
Monkeypox Virus I1L (strain Zaire-96-I-16), mFc Tag |
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E24PVA026 | EnoGene | 50 μg |
Monkeypox Virus A29L (strain Zaire-96-I-16), His Tag |
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E24PVA016 | EnoGene | 50 μg |
Monkeypox Virus A35R (strain Zaire-96-I-16), mFc Tag |
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E24PVA022 | EnoGene | 50 μg |
Monkeypox Virus B21R (strain Zaire-96-I-16), His Tag |
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E24PVA023 | EnoGene | 50 μg |
Monkeypox Virus A33R (strain Zaire-96-I-16), His Tag |
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E24PVA027 | EnoGene | 50 μg |
Monkeypox Virus A33R (strain Zaire-96-I-16), mFc Tag |
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E24PVA028 | EnoGene | 50 μg |
Monkeypox Virus B2R Recombinant Protein N-His Tag Lyophilized |
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IMPXVB2RRNHISLY100UG | Innovative research | each |
Monkeypox Virus B2R Recombinant Protein N-His Tag Lyophilized |
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IMPXVB2RRNHISLY1MG | Innovative research | each |
Monkeypox Virus A35R Recombinant Protein C-His Tag Lyophilized |
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IMPXVA35RRCHISLY100UG | Innovative research | each |
Monkeypox Virus A35R Recombinant Protein C-His Tag Lyophilized |
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IMPXVA35RRCHISLY1MG | Innovative research | each |
Monkeypox Virus C18L Recombinant Protein N-His Tag Lyophilized |
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IMPXVC18LRNHISLY100UG | Innovative research | each |
Monkeypox Virus C18L Recombinant Protein N-His Tag Lyophilized |
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IMPXVC18LRNHISLY1MG | Innovative research | each |
Monkeypox Virus C19L Recombinant Protein N-His Tag Lyophilized |
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IMPXVC19LRNHISLY100UG | Innovative research | each |
Monkeypox Virus C19L Recombinant Protein N-His Tag Lyophilized |
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IMPXVC19LRNHISLY1MG | Innovative research | each |
Monkeypox virus (strain Zaire-96-I-16) E8L Protein, His Tag |
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E8L-M52H3 | ACROBIOSYSTEMS | 50ug |
Monkeypox virus (strain Zaire-96-I-16) A29L Protein, His Tag |
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A2L-M52H3 | ACROBIOSYSTEMS | 100ug |
Monkeypox virus (strain Zaire-96-I-16) A35R Protein, His Tag |
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A3R-M52H3 | ACROBIOSYSTEMS | 100ug |
Recombinant Monkeypox virus Envelope protein A28 homolog(A30L),partial |
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CSB-BP848659MHV | Cusabio | 20μg |
Recombinant Monkeypox virus Envelope protein A28 homolog(A30L),partial |
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CSB-EP848659MHV-B | Cusabio | 20μg |
Recombinant Monkeypox virus Envelope protein A28 homolog(A30L),partial |
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CSB-MP848659MHV | Cusabio | 20μg |
Rabbit Anti Monkeypox Virus A30L Polyclonal Antigen Affinity Purified |
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IRBAMPXVA30LAAP100UG | Innovative research | each |
Rabbit Anti Monkeypox Virus B6R Polyclonal Antigen Affinity Purified |
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IRBAMPXVB6RAAP100UG | Innovative research | each |
Monkeypox Virus A35R Recombinant Protein N-GST and C-His Tag Lyophilized |
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IMPXVA35RRNGSTCHISLY100UG | Innovative research | each |
Monkeypox Virus A35R Recombinant Protein N-GST and C-His Tag Lyophilized |
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IMPXVA35RRNGSTCHISLY1MG | Innovative research | each |
Monkeypox virus (strain Zaire-96-I-16) L1R Protein, His Tag (HPLC verified) |
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L1R-M5241 | ACROBIOSYSTEMS | 500ug |
Monkeypox virus (strain Zaire-96-I-16) H3L Protein, His Tag (MALS verified) |
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H3L-M52H1 | ACROBIOSYSTEMS | 50ug |
Monkeypox virus (strain Zaire-96-I-16) A17L Protein, His Tag (HPLC verified) |
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A1L-M52H3 | ACROBIOSYSTEMS | 50ug |
Monkeypox virus (strain Zaire-96-I-16) A30L Protein, His Tag (MALS verified) |
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A3L-M5243 | ACROBIOSYSTEMS | 50ug |
Biotinylated Monkeypox virus (strain Zaire-96-I-16) E8L Protein, His,Avitag™ |
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E8L-M82E3 | ACROBIOSYSTEMS | 200ug |
Monkeypox Virus Zaire Strain 96-I-16 B5R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16B5RRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 B5R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16B5RRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 B6R Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16B6RRCHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 B6R Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16B6RRCHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 B6R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16B6RRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 B6R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16B6RRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 E8L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16E8LRCHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 E8L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16E8LRCHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 E8L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16E8LRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 E8L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16E8LRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 H3L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16H3LRCHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 H3L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16H3LRCHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 H3L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16H3LRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 H3L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16H3LRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 M1R Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16M1RRCHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 M1R Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16M1RRCHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 M1R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16M1RRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 M1R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16M1RRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A27L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A27LRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A27L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A27LRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A29L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16A29LRCHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A29L Recombinant Protein C-His Tag Lyophilized |
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IMPXVZ96I16A29LRCHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A29L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A29LRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A29L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A29LRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A33R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A33RRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A33R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A33RRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A36R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A36RRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 A36R Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16A36RRNHISLY1MG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 D13L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16D13LRNHISLY100UG | Innovative research | each |
Monkeypox Virus Zaire Strain 96-I-16 D13L Recombinant Protein N-His Tag Lyophilized |
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IMPXVZ96I16D13LRNHISLY1MG | Innovative research | each |
The clinical performance of the MPXV-RPA-assay was tested using 47 sera and whole blood samples from humans collected during the recent MPXV outbreak in Nigeria as well as 48 plasma samples from monkeys some of which were experimentally infected with MPXV. The specificity of the MPXV-RPA-assay was 100% (50/50), while the sensitivity was 95% (43/45). This new MPXV-RPA-assay is fast and can be easily utilised at low resource settings using a solar powered mobile suitcase laboratory.