Evaluation and clinical validation of monkeypox (mpox) virus real-time PCR assays.
Evaluation and clinical validation of monkeypox (mpox) virus real-time PCR assays.
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DOI：
10.1016/j.jcv.2022.105373
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发表时间：
2023-02
影响因子：
8.8
通讯作者：
Greninger, Alexander L.
中科院分区：
文献类型：
--
作者：
Mills, Margaret G.;Juergens, Kate B.;Gov, Jolene P.;McCormick, Carter J.;Sampoleo, Reigran;Kachikis, Alisa;Amory, John K.;Fang, Ferric C.;Perez-Osorio, Ailyn C.;Lieberman, Nicole A. P.;Greninger, Alexander L.
2022年春，猴痘（天花）疫情在全球范围内蔓延。在此，我们描述了猴痘病毒（MPXV）特异性和泛正痘病毒qPCR检测试剂盒用于临床的性能特征。我们验证了靶向MPXV特异性基因座F3 L和G2 R（分别为基因MPXVgp 052/OPG 065和MPXVgp 002和gp 190/OPG 002）的基于探针的qPCR测定法以及靶向E9 L基因座（MPXVgp 057/OPG 071）的泛正痘病毒测定法。使用Roche MP 96或Promega麦克斯韦48仪器提取临床样品和合成对照。在AB 7500热循环仪上进行qPCR。通过液滴PCR定量合成对照DNA和高浓度临床样品。评价了骆驼痘和牛痘基因组DNA、牛痘培养上清液以及HSV和VZV阳性临床标本的交叉反应性。我们还使用干拭子、Aptima阴道和直肠拭子、鼻咽、直肠和口腔拭子、脑脊液、血浆、血清、全血、母乳、尿液、唾液和精液检测了F3 L检测试剂盒的性能。在病毒转运培养基/通用转运培养基（VTM/UTM）中病毒DNA的检测限（LoD）为65.6拷贝/mL或3.3拷贝/PCR反应时，MPXV-F3 L检测试剂盒具有重现性。未观察到与疱疹病毒或其他痘病毒的交叉反应性。MPXV-F3 L可检测替代标本类型中的MPXV DNA，LoD范围为260-1000拷贝/mL或5.7-10拷贝/PCR反应。在临床拭子VTM标本中，MPXV-F3 L和MPXV-G2 R检测试剂的平均性能分别优于OPXV-E9 L 2.4和2.8 Ct。MPXV-G2 R比MPXV-F3 L高0.4Ct，这与MPXV基因组的不稳定反向末端重复序列（ITR）中存在两个拷贝的G2 R一致。使用三种临床验证的测定法通过qPCR容易地检测MPXV。
In spring of 2022, an outbreak of monkeypox (mpox) spread worldwide. Here, we describe performance characteristics of monkeypox virus (MPXV)-specific and pan-orthopoxvirus qPCR assays for clinical use. We validated probe-based qPCR assays targeting MPXV-specific loci F3L and G2R (genes MPXVgp052/OPG065 and MPXVgp002 and gp190/OPG002, respectively) and a pan-orthopoxvirus assay targeting the E9L locus (MPXVgp057/OPG071). Clinical samples and synthetic controls were extracted using the Roche MP96 or Promega Maxwell 48 instrument. qPCR was performed on the AB7500 thermocycler. Synthetic control DNA and high concentration clinical samples were quantified by droplet PCR. Cross-reactivity was evaluated for camelpox and cowpox genomic DNA, vaccinia culture supernatant, and HSV- and VZV-positive clinical specimens. We also tested the performance of the F3L assay using dry swabs, Aptima vaginal and rectal swabs, nasopharyngeal, rectal, and oral swabs, cerebrospinal fluid, plasma, serum, whole blood, breastmilk, urine, saliva, and semen. The MPXV-F3L assay is reproducible at a limit of detection (LoD) of 65.6 copies/mL of viral DNA in viral transport medium/universal transport medium (VTM/UTM), or 3.3 copies/PCR reaction. No cross-reactivity with herpesviruses or other poxviruses was observed. MPXV-F3L detects MPXV DNA in alternative specimen types, with an LoD ranging between 260-1000 copies/mL, or 5.7-10 copies/PCR reaction. In clinical swab VTM specimens, MPXV-F3L and MPXV-G2R assays outperformed OPXV-E9L by an average of 2.4 and 2.8 Cts, respectively. MPXV-G2R outperformed MPXV-F3L by 0.4 Cts, consistent with presence of two copies of G2R present in labile inverted terminal repeats (ITRs) of MPXV genome. MPXV is readily detected by qPCR using three clinically validated assays.
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影响因子：
158.5
作者：
Thornhill, John P.;Barkati, Sapha;Orkin, Chloe M.
通讯作者：
Orkin, Chloe M.
DOI：
10.3390/v13010043
发表时间：
2020-12-30
期刊：
Viruses
影响因子：
--
作者：
Silva NIO;de Oliveira JS;Kroon EG;Trindade GS;Drumond BP
通讯作者：
Drumond BP
DOI：
10.3390/v14112393
发表时间：
2022-10-29
期刊：
Viruses
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.1
作者：
Li, Yu;Zhao, Hui;Wilkins, Kimberly;Hughes, Christine;Damon, Inger K.
通讯作者：
Damon, Inger K.
影响因子：
11.8
作者：
McCollum, Andrea M.;Damon, Inger K.
通讯作者：
Damon, Inger K.