Mary Wilson
Well-known member
CORRESPONDENCE | ARTICLES IN PRESS
Available online 19 August 2020
In Press, Journal Pre-proof
Published:August 19, 2020
DOI:https://doi.org/10.1016/j.pathol.2020.08.002
Claire Y.T. Wang, Cameron Buckley, Cheryl Bletchly, Patrick Harris, David Whiley
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in Wuhan, China, in December 2019 as the aetiological agent of Coronavirus disease 2019 (COVID-19).[SUP]1[/SUP][SUP],[/SUP][SUP]2[/SUP] Since then, the disease has spread rapidly worldwide and the World Health Organization (WHO) declared a pandemic on 11 March 2020.[SUP]3[/SUP][SUP],[/SUP][SUP]4[/SUP] At the beginning of the outbreak, rapid development and implementation of reliable detection methods became an immediate priority for clinical laboratories worldwide, and RT-PCR methods, including those provided by the WHO,[SUP]5[/SUP][SUP],[/SUP][SUP]6 [/SUP]have been implemented broadly. At the early stage of the outbreak, however, positive control material for RT-PCR assays (from positive patient samples, or viral culture) were not readily available. In such circumstances laboratories often turn to using synthetic controls (synDNA fragments or plasmids).[SUP]7[/SUP][SUP],[/SUP][SUP]8[/SUP]
These synthetic controls have their advantages, particularly in that the controls can be acquired as readily as PCR primers and probes. Yet, depending on how they are designed, precautions must be taken when handling such controls as trace amounts of this material can potentially cause contamination in the same way as that caused by PCR products. Here we report contamination of a SARS-CoV-2 probe that our evidence suggests occurred at the oligonucleotide manufacturer, and was due to the manufacturer synthesising full length control oligonucleotides (spanning from the forward to reverse primers) in parallel with our probe orders.
https://www.pathologyjournal.rcpa.ed...885-0/fulltext
Available online 19 August 2020
In Press, Journal Pre-proof
Published:August 19, 2020
DOI:https://doi.org/10.1016/j.pathol.2020.08.002
Claire Y.T. Wang, Cameron Buckley, Cheryl Bletchly, Patrick Harris, David Whiley
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in Wuhan, China, in December 2019 as the aetiological agent of Coronavirus disease 2019 (COVID-19).[SUP]1[/SUP][SUP],[/SUP][SUP]2[/SUP] Since then, the disease has spread rapidly worldwide and the World Health Organization (WHO) declared a pandemic on 11 March 2020.[SUP]3[/SUP][SUP],[/SUP][SUP]4[/SUP] At the beginning of the outbreak, rapid development and implementation of reliable detection methods became an immediate priority for clinical laboratories worldwide, and RT-PCR methods, including those provided by the WHO,[SUP]5[/SUP][SUP],[/SUP][SUP]6 [/SUP]have been implemented broadly. At the early stage of the outbreak, however, positive control material for RT-PCR assays (from positive patient samples, or viral culture) were not readily available. In such circumstances laboratories often turn to using synthetic controls (synDNA fragments or plasmids).[SUP]7[/SUP][SUP],[/SUP][SUP]8[/SUP]
These synthetic controls have their advantages, particularly in that the controls can be acquired as readily as PCR primers and probes. Yet, depending on how they are designed, precautions must be taken when handling such controls as trace amounts of this material can potentially cause contamination in the same way as that caused by PCR products. Here we report contamination of a SARS-CoV-2 probe that our evidence suggests occurred at the oligonucleotide manufacturer, and was due to the manufacturer synthesising full length control oligonucleotides (spanning from the forward to reverse primers) in parallel with our probe orders.
https://www.pathologyjournal.rcpa.ed...885-0/fulltext