CSI: Rodent, brilliant! The MERS-CoV RNA was spiked in the total RNA extracted from the swabs to evaluate the assay panel. But you do have to be able to look at data and tell a story. Ten-fold dilutions of the viral RNA (ranging from 1.2 × 10−1 to 1.2 × 106 copies/μl) were also used to evaluate the two RT-RPA-VF assays under the same reaction conditions. And sometimes Access. I understand if you keep that side of you under wraps around normal people. The gold-labeled anti-FITC antibody and streptavidin fixed on the test paper will capture the amplicon labeled with FITC and biotin at both ends and accumulate to form a red band. Selon la commission des comptes de la sécurité sociale Rapport septembre 2020 volume 1 p. This seems to be under represented in “how to be a data scientist” posts but it is very important. Clearly there are very rigorous requirements for a proper data scientist. Considering recent experience related to the prevention and control of COVID-19, a rapid diagnostic assay is important in the initial emergency response stage of an epidemic to reduce the workload of laboratory technicians, reduce the risk of cross-infection and ensure the timely treatment of patients. Tell me a story. According to previous reports, fluid samples can be quickly heated directly to release RNA without RNA purification and used in the RT-RPA assay [37,38]. A pocos meses de finalizar el 2019, son muy pocas las entidades que conocen sobre la nueva interpretación a las Normas Internacionales de Información Financiera, CINIIF 23, y menos aún que su aplicación ya es obligatoria para todas aquellas entidades que emiten sus estados financieros bajo Normas Internacionales de Información Financiera (NIIF’s). ... Reddit veut lever plus de 500 millions de dollars. Then, 2.5 μl of 280 mM MgCl2 were added to the tube cap. Meanwhile, the THF site of probe was cleaved by endonuclease IV (Nfo), thereby removing the blocking group and forming a new 3’ end that can be extended [21]. At the end of the day, I'm not sure that I'm a prototypical data scientist, but I think that few people who do "data science" (whatever that means) are. One final note: most companies (especially places like Facebook, Google and LinkedIn) do strong culture screens. Writing – original draft, Affiliations Writing – review & editing, * E-mail: yuwei0901@outlook.com (YG); xiaxzh@cae.cn (XX); wanghualei@jlu.edu.cn (HW). So convince me that you can write code. The UpE RNA transcript and N RNA transcript were subjected to 10-fold serial dilutions. Yes What application(s) that can store and display data is almost guaranteed to be on every clients desktop and they all know how to use? Therefore, the sensitivity of each RT-RPA-VF assay was sufficient for a MERS-CoV diagnosis. However, the RT-RPA-VF assay targeting the N gene detected as low as 1.2 copies/μl viral RNA, with a sensitivity equivalent to the RT-qPCR assay for the N gene. Marketing Insider Group mentions several ways social media can help advertise business products and services. Based on these results, the RT-RPA-VF assay panel has the potential to be used as an alternative to RT-qPCR for MERS-CoV. Specificity testing was conducted in a Biosafety Level 2 facility to ensure adequate biosafety in the laboratory. Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun, China, Affiliation You need to understand data. Yes First, 10-fold dilutions of synthesized RNA transcripts were used as a template for the RT-RPA-VF assays, and the reaction process was performed at different temperatures (42°C, 40°C, 39°C, 37°C, 35°C, 33°C, and 30°C) for 30 min. Other times I'm in a room full of highly successful but data illiteral individuals who don't care how it works or why. I’ve noticed a bunch of posts here that are of the form “I’m a X major and want to be a data scientist, what do I do?” so I thought I would chime in and give my opinion. In our assay panel, the limit of detection of the viral RNA was 1.2 × 101 copies/μl for the UpE assay and 1.2 copies/μl for the N assay in 30 min. The squirrel hypothetical is my new go to interview question! Ten-fold dilutions of viral RNA were used as templates for the two RT-RPA-VF assays under the same reaction conditions to further validate the sensitivity. College of Life Sciences, Shandong Normal University, Jinan, China, Affiliation A lot of people need to get off the whole machine learning hype train and do what is best for the business, using what tools are best for the business. Digital currencies are the payment methods for the future. Hopefully it helps. Compared with gold standard RT-qPCR assays, the sensitivity of the RT-RPA-VF assay for the UpE gene was less than the 1.2 copies/μl detection limit of the RT-qPCR for the UpE gene. Do My Homework sagt am Jul 26, 2019 @ 08:14: Social media marketing isn’t a new phenomenon. You need imagination. Compared with PCR technology, the RT-RPA-VF assay has eliminated the need for precision temperature cycling instruments [15]. College of Veterinary Medicine, South China Agricultural University, Guangzhou, China, Affiliations In addition, a closed vertical flow visualization strip (VF) is a terminal detection device for detecting RT-RPA products, and this design is particularly applicable in low-resource settings. https://doi.org/10.1371/journal.pntd.0009227.t007. With the right channel and the right content, a business could capitalize on a particular market relatively quickly. https://doi.org/10.1371/journal.pntd.0009227.g001. The World Health Organization (WHO) recommends real-time RT-PCR targeting the sequence upstream of the envelope protein gene (UpE) and the open reading frame 1ab (ORF1ab) or the gene encoding the nucleocapsid protein gene (N) as the screening and diagnostic targets for MERS-CoV [10–12]. For that you’ll need PERL, Python, VB, etc. The amplification reaction was performed using an Mx3005P real-time PCR instrument (Agilent Company, California, USA), and thermal cycling was performed at 45°C for 5 min, followed by 94°C for 30 s and 40 cycles of 94°C for 5 s and 60°C for 30 s. Each group included a negative control. More remarkably, SARS-CoV-2 has infected more than six million people in the last six months. If you see a dead squirrel on the top floor of a parking garage and your first thought is “Ewwwww, a dead squirrel” I don’t want you. According to the highest dilution at which 100% of replicates were positive, the optimal reaction time for the two assays was 25 min, as shown in Table 5. https://doi.org/10.1371/journal.pntd.0009227.t005. The mean viral load in the lower respiratory tract of MERS-CoV infected patients is approximately 5.01 × 106 copies/ml [32]. Fiche de paie Excel pour 2021 est téléchargeable sur votre site bulletinspaie.com, dés le 05/01/2021 , les taux et les nouveaux barèmes 2020 sont également disponible . It would be awesome if you had minions to do the shit work for you but let’s face it, if you are just coming out of school you are the minion. Additionally, its primers and probe are relatively complicated. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China, Affiliations The coincidence rate between RT-RPA-VF and RT-qPCR for the UpE gene at 87.5% (kappa = 0.6). Methodology, Therefore, 33°C and 35°C were deemed the optimal temperatures for the RT-RPA-VF assays to detect the UpE and N genes, respectively. After mixing and centrifugation, the tubes were placed in a small metal bath at 33°C (UpE assay) or 35°C (N assay) for 25 min for amplification. The limit of detection of the RT-RPA-VF assay targeting the N gene was 1.2 copies/μl viral RNA, with a sensitivity equivalent to the RT-qPCR assay (Fig 4B). We typically turn people down, not because they can't hack the job, but because they don't fit with the company culture--and we're a small company. New comments cannot be posted and votes cannot be cast, More posts from the datascience community. Many times I wished I had a VB programmer so I could make what I delivered a lot slicker than a dump of shit into a spreadsheet. Read this far and realized you deserve a medal. On the other hand, the two RT-RPA-VF assays targeting the MERS-CoV UpE and N genes were rapid (within 30 min from amplification to observation) and highly sensitive (as little as 1.2–1.2 × 101 copies/μl viral RNA), and no sophisticated equipment was needed. Because dromedary camels have a large geographical range, the disease is subclinical and camels are significant animal products, vaccination and culling to block animal transmission are difficult to implement [34–36]. Finally, you need an area of technical analytic expertise. Approximately 84% of human cases have been reported in the tropical region of Saudi Arabia. All primers and probes were synthesized by Bao Biological, Co., Ltd. (Dalian, China). We speculate that this assay will be combined with quickly handled fluid samples without RNA extraction, showing potential for use in point-of-care testing for highly contagious tropical diseases. They just want to know how all of those fancy looking charts on the screen turns into $$$$$. Thank you for the blog. Roles At the same time, an SSB binds to the non-template strand to prevent the dissociation of the primer. The specificity of the RT-RPA-VF assays against the UpE (A) and N (B) genes. If you use the term “p-value” while explaining results to a client I’ll dock your pay. Conceptualization, MERS-CoV is a zoonotic pathogen, and the dromedary camel is the main host for transmission to humans. We thank all participators who work in Seven Peak Alpaca Farm and Northeast Camel Farm for their support with obtaining respiratory swabs. The two assays exhibited no cross-reactivity with multiple CoVs, including the bat severe acute respiratory syndrome related coronavirus (SARSr-CoV), the bat coronavirus HKU4, and the human coronaviruses 229E, OC43, HKU1 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Just keep asking yourself if what you are investigating will ultimately be useful to the client. A risk of another outbreak exists after largest outbreak occurred in the Republic of Korea, Saudi Arabia and the United Arab Emirates because MERS-CoV can be transmitted by two main routes: dromedary camels-to-humans and human-to-human [8,9]. This is where your background in stats, machine learning, natural language processing, etc. As some governments accept the digital currency as a mode of payment, we feel it is important for you to know some of these digital currencies and how they operate. Real-time RT-PCR has great advantages in sensitivity and specificity compared with other molecular techniques; nevertheless, it is only suitable for well-equipped or central laboratories due to the demand for sophisticated alternating temperature and fluorescent capture instruments. Ten-fold dilutions of synthesized UpE RNA transcripts were used to establish the standard curve, and 100-fold dilutions of total RNA from virus-infected cell suspensions were used as the test template. A place for data science practitioners and professionals to discuss and debate data science career questions. Subsequently, the probe annealed to the single-stranded target sequence within the amplification product through two primer extension. If you have to learn how to use a test tube, and you probably will, then do it. And they were completely obtuse. Take A Sneak Peak At The Movies Coming Out This Week (8/12) Olivia Munn using her platform stepping up to #StopAsianHate; Travel through Daylight Savings Time with these 16 time travel movies The u/AutoModerator community on Reddit. The conserved sequences of the UpE and N genes of the EMC strain were used as templates to design primers and probes. As liquid flows on the strip, amplicons labeled with biotin can bind to gold particles to form a complex because biotin can bind to streptavidin. Sooner or later you will need to impose structure on data or the data that is given to you will be highly structured. This approach results in a significant delay in the reporting of test results, which delays epidemic outbreak control and treatment. And the candidates seem much more qualified after a few shots. Total RNA extracted from swab samples that was not spiked with the MERS-CoV RNA was used as a control group. I don’t want those people. You don't have to be proficient in everything, because even I spend a fair amount of time on stack overflow. Clearly there are very rigorous requirements for a proper data scientist, much of which cannot be taught in a classroom, so it seems like the best way to actually become a data scientist is to gain some experience, leaving us in a catch-22 situation. The results showed that swab RNA did not affect the limit detection and specificity of the RT-RPA-VF assays. Thus, the four enzymes of RT-RPA have strong activity within a certain temperature range, but the binding and extension of the primers and probe to the template must occur at a specific temperature to maximize their effectiveness. Citation: Huang P, Jin H, Zhao Y, Li E, Yan F, Chi H, et al. People who work in camel farms, slaughterhouses and markets are at risk, and infected people may spread the virus to their contacts. My first results that I delivered to a client many years ago were horribly wrong because of some fundamentally incorrect assumptions I made about the completeness of the data that I would have known about had I asked. First, specific primers and probes were designed; the probe was labeled with fluorescein isothiocyanate (FITC) at the 5′ end and it contained a tetrahydrofuran (THF) residue that replaced a nucleotide and a blocking group C3-spacer at the 3′ end. At the end of January 2020, 2519 laboratory confirmed cases with a case-fatality rate of 34.3% have been reported. You need to be able to convert your brilliant analysis into something that normal people can understand. As a statistician who recently transitioned into a more prototypical data scientist role, all I can say is that you really have to know your audience. No, Is the Subject Area "SARS CoV 2" applicable to this article? https://doi.org/10.1371/journal.pntd.0009227.g003. The design of the RT-RPA-VF assay can also be used to develop methods for the rapid detection of other infectious diseases. https://doi.org/10.1371/journal.pntd.0009227.t001. Thus, this panel may be preferred to rapidly detect MERS-CoV in low-resource settings, even as a point-of-care approach for the timely prevention and control of a pandemic. Next, the concentration of the MERS-CoV RNA was calculated as 1.2 × 109 copies/μl in 461 ng/μl total RNA or 5 × 106 PFU/ml virus in using a positive correlation between cycles and concentration. Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China, Affiliation My friends and close relatives all think I’m strange when I start wondering out loud about things like that. Unlike SARS-CoV, which was brought under control within one year after the outbreak, MERS continues to infect people. I totally get your point about hacking the data though. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China, The complete sequences of 11 representative MERS-CoV strains from GenBank reported from 2012 to 2015 were aligned, and the conserved sequences of the UpE and N genes were screened as targets. Compared to other MERS-CoV molecular detection methods, such as PCR, nested PCR, RT-qPCR, and the one-pot RT-LAMP assay, the RT-RPA-VF assay provides a shorter reaction time at a lower temperature [12,29,30]. Chemistry is about understanding the world at distances from an angstrom to a micron (ish). Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun, China, Affiliation You will also need to interact with clients on a continual basis since you may understand analytic techniques but you most likely won’t understand the domain you are working in. Detection of 10-fold serial dilutions of the MERS-CoV RNA using the RT-RPA-VF assays against the UpE (A) and N (B) genes. Therefore, the rapid diagnosis of MERS-CoV is the key to successful containment and prevention of its spread. Compared with antigen detection methods, such as enzyme-linked immunosorbent assays and colloidal gold immunochromatographic assay, nucleic acid amplification detection methods have the disadvantages of requiring professional technical operators and sophisticated equipment, but they have higher sensitivity and accuracy. And if you don't fit in, you don't fit in---that's ok. More important than anything is being business savvy and actually knowing what your stakeholders want and how to achieve this. Share photos and videos, send messages and get updates. An absolute RT-qPCR assay was performed using primer and probe sequences against the UpE gene described in a previous report to quantify the concentration of the MERS-CoV RNA in a total RNA sample containing 5 × 106 PFU/ml virus in the culture [11]. A variety of MERS-CoV detection methods have been established, including nucleic acid amplification, antigen detection, and serological detection. I do as well. You need to explain your results, why they are significant and why someone should trust them in a way that civilians understand. Real-time RT-PCR has great advantages in terms of high sensitivity and technological maturity; however, the need for sophisticated instruments has led to the delivery of specimens from low-resource settings to highly equipped centralized laboratories for testing. In addition, the reverse amplification primer was labeled with biotin at the 5′ end. RNA transcripts of the partial UpE (27240–27589) and N (29185–29515) genes of the MERS-CoV EMC strain (GenBank No. Finally, the amplicons were indirectly labeled with FITC and biotin due to the extension of the probe and reverse amplification primer [22]. Go fork some repo and commit to it (I know for a fact that twitter hires data scientists from people who commit to certain repos). Make sure you have some analytics on your resume, and that you can do stuff with Python and/or R, and that you know some SQL. Ten-fold dilutions of synthesized UpE-RNA transcripts were used to establish the standard curve, 100-fold dilutions of total RNA from virus-infected cell suspensions were used as the test template, and the RNA of uninfected cell suspensions was used as the negative control template. Furthermore, this assay can minimize risk of false positives and the requirement to ship specimens. Thanks for the helpful post. This panel displays good sensitivity and specificity and may be useful to rapidly detect MERS-CoV early during an outbreak and for disease surveillance. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. And I’m more interested in hacking skills more than formal systems development. This is a great inspiring article. Unless you are lucky enough to have a client that understands all that stuff. And when not to? Symptoms of MERS are atypical, including fever, cough, shortness of breath and occasionally pneumonia and gastrointestinal symptoms.