The sequence-independent, single-primer amplification (SISPA) enables the random amplification of nucleic acids, allowing the detection and genome sequencing of different viral agents. This feature of SISPA method provides evidence for application of it in monitoring the presence of adventitious RNA viruses in cell cultures. We evaluated SISPA method for the detection of a challenge RNA virus representing adventitious agent in cell cultures. Besides, by optimizing the SISPA method in our laboratory, we found false-positive results on negative control lanes in electrophoresis gels. To investigate the sources of contamination, false-positive results of SISPA were cloned into Escherichia coli cells, sequenced, and phylogenetically analyzed. This data revealed that the SISPA method can be used as an adjunct method to confirm the absence of unexpected adventitious RNA viruses in cell cultures. The phylogenetic analysis of SISPA contaminant sequences showed that the false-positive results were caused by nucleic acid amplification of commercial cDNA synthesis kit reagents, probably tracing back to expression plasmids and host ribosomal sequences, used for the production of enzymes. Therefore, laboratories using random amplification methods must be constantly aware of the potentials of such contaminations, yielding false-positive results and background noise in the final NGS reads.
Publishing with the International Journal of Clinical and Experimental Ophthalmology was a rewarding experience as review process was thorough and brisk. Their visibility online is second to none as t...
University of Port Harcourt Teaching Hospital, Nig...
Dr. Elizabeth A Awoyesuku
Really good service with prompt response. Looking forward to having long lasting relationship with your journal
Avishek Bagchi
I like the quality of the print & overall service. The paper looks quite impressive. Hope this will attract interested readers. All of you have our best wishes for continued success.
Arshad Khan
“It was a delightful experience publishing my manuscript with the Clinical Journal of Obstetrics and Gynecology. They offered me lots of opportunities I never had from most publishing houses and the...
Asafo Jones
Thank you for your attitude and support. I am sincerely grateful to you and the entire staff of the magazine for the high professionalism and fast quality work. Thank you very much!
USA
Igor Klepikov
Publishing an article is a long process, but working with your publication department made things go smoothly, even though the process took exactly 5 months from the time of submitting the article til...
Anas Diab
“The choice to submit the forensic case study to the Journal of Addiction Therapy and Research was dictated by the match between the content and the potential readership. The publication process pro...
Ph.D, Boston University Department of Communicatio...
Elisabeth H. Wiig
Regarding to be services, we note that are work with high standards of professionalism translated into quick response, efficiency which makes communication accessible. Furthermore, I believe to be muc...
Amélia João Alice Nkutxi
We really appreciate and thanks the full waiver you provide for our article. We happy to publish our paper in your journal. Thank you very much for your good support and services.
Ali Abusafia
It was a real pleasure working with your team. The review was done fast, and it was very clear, the editing was flawless, the article was published quickly compared to other journals, and everyone w...
HSPI: We're glad you're here. Please click "create a new Query" if you are a new visitor to our website and need further information from us.
If you are already a member of our network and need to keep track of any developments regarding a question you have already submitted, click "take me to my Query."