Validation of a pan-orthopox real-time PCR assay for the detection and quantification of viral genomes from nonhuman primate blood

Mucker, Eric M.; Hartmann, Christopher J.; Hering, Donna; Giles, Wendy; Miller, David; Fisher, Robert W.; Huggins, John W.

doi:10.1186/s12985-017-0880-8

Cited by 10 publications

(10 citation statements)

References 10 publications

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“…DNA extraction and PCR analysis methodology DNA extraction with QIAamp DNA Mini Kit (Qiagen), inactivation of infectious virus by incubation of specimen in extraction buffer at 56° C for 1 hr, and pan-orthopox real time polymerase chain reaction (PCR) assays were performed using a Roche LightCycler and software (24). All reagents were aliquoted into single test aliquots for up to 8 samples plus controls at USAMRIID.…”

Section: Clinical Laboratory Proceduresmentioning

confidence: 99%

Clinical characterization of human monkeypox infections in the Democratic Republic of the Congo

Pittman

Martin

Placide

et al. 2022

Preprint

109

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We describe the results of a prospective observational study of the clinical natural history of human monkeypox virus (MPXV) infections at the remote General Hospital in Kole, DRC (Kole hospital), the rainforest of the Congo River basin of the Democratic Republic of the Congo (DRC) from March 2007 until August 2011. The research was conducted jointly by the Institute National de Recherche Biomedical (INRB) and the US Army Medical Research Institute of Infectious Diseases (USAMRIID). The Kole hospital was one of the two previous WHO Monkeypox (MPX) study sites (1981-1986). The hospital is staffed by the Missionaries of Christ Jesus, a Spanish Order of Catholic Sisters who have continued the treatment of patients with a clinical diagnosis of MPXV infection. Of 244 patients admitted with a clinical diagnosis of MPXV infection, 216 were positive in both the Pan-Orthopox and MPX specific PCR. The cardinal observations of these 216 patients are summarized in this report. There were three deaths (3/216) among these hospitalized patients; fetal death occurred in 4 of 5 (80%) patients who were pregnant at admission. The most common complaints were rash (96.8%), malaise (85.2%), sore throat (78.2%), and lymphadenopathy/adenopathy (57.4%). The most common physical exam findings were MPX rash (99.5%) and lymphadenopathy (98.6%). Age group of less than 5 years had the highest lesion count. Primary household cases tended to have higher lesion counts than secondary or later same household cases. Of the 216 patients, 200 were tested for IgM & IgG antibodies (Abs) to Orthopoxviruses. All 200 patients had anti-orthopoxvirus IgG Abs; whereas 189/200 were positive for IgM. Patients with hypoalbuminemia had a high risk of severe disease. Patients with fatal disease had significantly higher maximum geometric mean values than survivors for the following variables, respectively: viral DNA in blood (DNAemia, p=0.0072); maximum lesion count (p=0.0025); day of admission mean AST and ALT (p=0.0002 and p = 0.0224, respectively, adjusted p-values).

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Section: Clinical Laboratory Proceduresmentioning

confidence: 99%

Clinical characterization of human monkeypox infections in the Democratic Republic of the Congo

Pittman

Martin

Placide

et al. 2022

Preprint

109

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“…More striking is the high ranking of Russia. After the eradication of SPX in 1980, the agent of variola virus was held captive in a Russian (VECTOR: State Research Center of Virology and Biotechnology ) and a US laboratory [ 39 ], which explains Russia's comparatively high and constant involvement in mpox research. China’s relatively low rank in mpox publications is also striking.…”

Section: Discussionmentioning

confidence: 99%

Global mpox research in the light of the current outbreak: demands, drivers, and obstacles

Klingelhöfer

Braun

Groneberg

et al. 2023

Emerging Microbes & Infections

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Following the current outbreak, the mpox virus (formerly: monkeypox virus) is a highly threatening pathogen with public health significance, although mpox is still considered a neglected disease. Previously confined mainly to Africa, the virus spread globally in 2022. However, knowledge about mpox is limited, causing a distorted perception of the disease. Therefore, this study aimed to collect all information on scientific mpox publishing and to analyse them according to their chronological, geographical, and epidemiological patterns. It was not until the global outbreak that the relatively small number of publications was replaced by the immense increase in annual publication numbers. The most important player is the USA with a central role in international networking. They collaborated mainly with the Democratic Republic of Congo, a primary endemic country where the first viral clades were determined. Nigeria and other African countries were also represented, although mainly in the form of co-authorships. The fact that few of the first authors are from low- or middle-economic countries demonstrates the need to promote equitable networking at the global level and their support for surveillance and targeted immunization programmes.

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“…DNA extraction with QIAamp DNA Mini Kit (Qiagen), inactivation of infectious virus by incubation of specimen in extraction buffer at 56° C for 1 hour, and pan-orthopox real time polymerase chain reaction (PCR) assays were performed using a Roche LightCycler and software [ 27 ]. All reagents were aliquoted into single test aliquots for up to 8 samples plus controls at USAMRIID.…”

Section: Methodsmentioning

confidence: 99%

Clinical characterization and placental pathology of mpox infection in hospitalized patients in the Democratic Republic of the Congo

et al. 2023

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We describe the results of a prospective observational study of the clinical natural history of human monkeypox (mpox) virus (MPXV) infections at the remote L’Hopital General de Reference de Kole (Kole hospital), the rainforest of the Congo River basin of the Democratic Republic of the Congo (DRC) from March 2007 until August 2011. The research was conducted jointly by the Institute National de Recherche Biomedical (INRB) and the US Army Medical Research Institute of Infectious Diseases (USAMRIID). The Kole hospital was one of the two previous WHO Mpox study sites (1981–1986). The hospital is staffed by a Spanish Order of Catholic Nuns from La Congregation Des Seours Missionnaires Du Christ Jesus including two Spanish physicians, who were members of the Order as well, were part of the WHO study on human mpox. Of 244 patients admitted with a clinical diagnosis of MPXV infection, 216 were positive in both the Pan-Orthopox and MPXV specific PCR. The cardinal observations of these 216 patients are summarized in this report. There were three deaths (3/216) among these hospitalized patients; fetal death occurred in 3 of 4 patients who were pregnant at admission, with the placenta of one fetus demonstrating prominent MPXV infection of the chorionic villi. The most common complaints were rash (96.8%), malaise (85.2%), sore throat (78.2%), and lymphadenopathy/adenopathy (57.4%). The most common physical exam findings were mpox rash (99.5%) and lymphadenopathy (98.6%). The single patient without the classic mpox rash had been previously vaccinated against smallpox. Age group of less than 5 years had the highest lesion count. Primary household cases tended to have higher lesion counts than secondary or later same household cases. Of the 216 patients, 200 were tested for IgM & IgG antibodies (Abs) to Orthopoxviruses. All 200 patients had anti-orthopoxvirus IgG Abs; whereas 189/200 were positive for IgM. Patients with hypoalbuminemia had a high risk of severe disease. Patients with fatal disease had higher maximum geometric mean values than survivors for the following variables, respectively: viral DNA in blood (DNAemia); maximum lesion count; day of admission mean AST and ALT.

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Validation of a pan-orthopox real-time PCR assay for the detection and quantification of viral genomes from nonhuman primate blood

Cited by 10 publications

References 10 publications

Clinical characterization of human monkeypox infections in the Democratic Republic of the Congo

Clinical characterization of human monkeypox infections in the Democratic Republic of the Congo

Global mpox research in the light of the current outbreak: demands, drivers, and obstacles

Clinical characterization and placental pathology of mpox infection in hospitalized patients in the Democratic Republic of the Congo

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