Validation Study of a Direct Real-Time PCR Protocol for Detection of Monkeypox Virus

Chelsky, Zachary L.; Dittmann, David; Blanke, Timothy; Chang, Michael; Vormittag-Nocito, Erica; Jennings, Lawrence J.

doi:10.1016/j.jmoldx.2022.09.001

Cited by 16 publications

(10 citation statements)

References 12 publications

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“…Polymerase chain reaction (PCR) using swabs of skin lesions (vesicles, ulcers, and scabs) recommended for confirming infection in symptomatic individuals is used to detect MPXV [ 17 ]. In addition, MPXV DNA has been detected by PCR in a wide variety of samples such as throat, nasopharynx, blood, urine, saliva, and semen [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Detection of Monkeypox Virus according to The Collection Site of Samples from Confirmed Cases: A Systematic Review

et al. 2022

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Due to the rapid evolution of the monkeypox virus, the means by which the monkeypox virus is spread is subject to change. Therefore, the present study aims to analyze the detection of the monkeypox virus according to the collection site of samples from confirmed monkeypox cases. A systematic literature review was performed using PubMed, Scopus, Web of Science, and Embase databases until 5 October 2022. A total of 1022 articles were retrieved using the search strategy. After removing duplicates (n = 566) and examining by title, abstract, and full text, 65 studies reporting monkeypox case reports were included with a detailed description of risk factors, sexually transmitted infections (STIs), site of monkeypox virus-positive specimens, location of skin lesions, and diagnostic test. A total of 4537 confirmed monkeypox cases have been reported, of which 98.72% of the cases were male with a mean age of 36 years, 95.72% had a sexual behavior of being men who have sex with men, and 28.1% had human immunodeficiency virus (HIV). The most frequent locations of lesions in patients diagnosed with monkeypox were: 42.85% on the genitalia and 37.1% in the perianal region. All confirmed monkeypox cases were diagnosed by reverse transcriptase polymerase chain reaction (RT-PCR), and the most frequent locations of samples collected for diagnosis that tested positive for monkeypox virus were: 91.85% from skin lesions, 20.81% from the oropharynx, 3.19% from blood, and 2.43% from seminal fluid. The disease course of the cases with monkeypox was asynchronous, with no severe complications, and most patients did not report specific treatment but simply followed a symptomatic treatment.

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Section: Introductionmentioning

confidence: 99%

Detection of Monkeypox Virus according to The Collection Site of Samples from Confirmed Cases: A Systematic Review

et al. 2022

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“…Currently, MPXV testing relies on methods including polymerase chain reaction (PCR), 14,15 enzyme-linked immunosorbent assay (ELISA), 16 loop-mediated isothermal amplication (LAMP), 17 and microarray. 18 PCR is widely recognized as the gold standard for diagnosing various diseases, including monkeypox, due to its high accuracy and sensitivity.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Zhao

Wang

et al. 2023

Anal. Methods

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“…The main method for laboratory diagnosis of mpox is the quantitative polymerase chain reaction (qPCR) assay [ 22 , 23 , 24 , 25 ]. Due to its high sensitivity, specificity, speed and possibility for high-throughput screening, it is recommended by the WHO as a gold-standard diagnostic method [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a Novel Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Monkeypox Virus Infections

Zuo

Miao

et al. 2022

Viruses

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A recent outbreak of monkeypox virus (mpox) has prompted researchers to explore diagnostics as a means of impeding transmission and further spread. Rapid, sensitive, and specific methods are crucial for accurately diagnosing mpox infections. Here, we developed a loop-mediated isothermal amplification (LAMP) assay for the specific detection of mpox. The primer sets were designed to target regions in and around the N4R gene, and results showed a detection limit of 2 × 100 DNA copies, which is comparable to the gold-standard qPCR method currently used to detect mpox. Particularly, the assay provides results visible to the naked eye within 30 min. This test specifically detects mpox DNA with no cross-reactivity to related DNA viruses including Varicella Zoster Virus (VZV), Hepatitis B virus (HBV), Vaccinia virus (VACV), Herpes simplex virus-1 (HSV-1), Herpes simplex virus-2 (HSV-2), Human papillomavirus-16 (HPV-16) and Human papillomavirus-18 (HPV-18). Furthermore, the LAMP assay has been evaluated using clinical samples from laboratory-confirmed mpox patients and found to be consistent with the qPCR results. Our results show that this single-tube LAMP method can contribute to diagnosis of suspected mpox infections in the field and clinic, especially in regions with limited laboratory resources.

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Validation Study of a Direct Real-Time PCR Protocol for Detection of Monkeypox Virus

Cited by 16 publications

References 12 publications

Detection of Monkeypox Virus according to The Collection Site of Samples from Confirmed Cases: A Systematic Review

Detection of Monkeypox Virus according to The Collection Site of Samples from Confirmed Cases: A Systematic Review

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Development of a Novel Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Monkeypox Virus Infections

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