Biomedical Science - Questions regarding on the ELISA test | Science Help

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Hi, Smunga Was wondering if you are familiar with the ELISA test, i do have the worksheet which i can show you, and you can let me know what you think about this?
User_39030 | Sep 20, 2021, 15:57 PM
Sorry for the rate response. Had stepped out .I can work on it and kindly provide the work sheet .
smunga | Sep 20, 2021, 15:59 PM
are you there friend?
smunga | Sep 20, 2021, 16:01 PM
Yes, Hi ive attached the file let me know what you think?
User_39030 | Sep 20, 2021, 16:05 PM
I accidentally attached two of the same files, sorry about that
User_39030 | Sep 20, 2021, 16:05 PM
Hi going through the file.
smunga | Sep 20, 2021, 16:06 PM
are you also able to write in the explanation of the answers as I also want to learn how the answers have come about? sort of like a tutoring lesson?
User_39030 | Sep 20, 2021, 16:07 PM
I can handle it .But its abit involving but i can handle the task and i can work on it .Kindly assign so that i may start working on it
smunga | Sep 20, 2021, 16:09 PM
ok i will present the explanation. and also submit a turnitin report
smunga | Sep 20, 2021, 16:10 PM
Hello you can assign the task. Thank you
smunga | Sep 20, 2021, 16:14 PM
HI, sorry the messages are going through a bit slow. Thanks
User_39030 | Sep 20, 2021, 16:18 PM
I will assign you now, give me a moment please :)
User_39030 | Sep 20, 2021, 16:26 PM
hello friend am patiently waiting for the go ahead to start the task.
smunga | Sep 20, 2021, 16:27 PM
let me start on the task .Thanks dear
smunga | Sep 20, 2021, 16:30 PM
Thanks very much, You are welcome to just answer the questions, instead of doing it in a report style :) Let me know if you need help or any more information.
User_39030 | Sep 20, 2021, 16:31 PM
Thanks for making it easier .Should i include the references? since i should not write it as a report
smunga | Sep 20, 2021, 16:34 PM
You can definitely include references for me if that's ok, thanks
User_39030 | Sep 20, 2021, 16:37 PM
That okay with me rest assured quality work will be deliverd
smunga | Sep 20, 2021, 16:38 PM
OH, give me a moment, i forgot to provide you the data
User_39030 | Sep 20, 2021, 16:39 PM
was actually typing asking of thesame(lol)
smunga | Sep 20, 2021, 16:40 PM
I attached the practical 6 results as well as some extra info which is in PowerPoint explaining things in further detail for your own understanding?, i think the practical 7 results are already in the manual, i actually have no idea, so was hoping someone would go through it with me haha, but considering you will be explaining a little bit to me, that would help a lot
User_39030 | Sep 20, 2021, 16:53 PM
ok is there any other material that would aid in completing the task more smoothly?
smunga | Sep 20, 2021, 16:56 PM
The powerpoint for practical 6, has the ELISA results and ive also attached a INDIRECT ELISA results thats a excel file
User_39030 | Sep 20, 2021, 16:56 PM
for some reason I cant upload the practical 7 PowerPoint, is there any other way I could send it to you?
User_39030 | Sep 20, 2021, 16:57 PM
that will be a bit hard since all communication are done on the site. Kindly try again .
smunga | Sep 20, 2021, 16:59 PM
Its still loading, once its done I'll let you know, though the results are in the worksheet i have given you for prac 7 :)
User_39030 | Sep 20, 2021, 17:08 PM
OH its been loaded let me know if you're able to open the last file
User_39030 | Sep 20, 2021, 17:08 PM
it has opened on my end. You have been amazing thanks am now good to go. Will start Asap
smunga | Sep 20, 2021, 17:12 PM
Thank you so much! let me know if there's anything else i can do on my end !
User_39030 | Sep 20, 2021, 17:16 PM
well there is one thing you can do. Relax now you are in safe hands. I will submit the task within 24 hours max . so that you can go through it and we can adjust where you see fit. Hope that okay with you
smunga | Sep 20, 2021, 17:19 PM
YES! perfect!
User_39030 | Sep 20, 2021, 17:20 PM
I've uploaded two other files in regards to the abstract, a marking rubric just for your guidelines, and more info on how to do the abstract, feel free to take your time. Thank you:)
User_39030 | Sep 20, 2021, 17:26 PM
Hello friend i have completed the assignment. Kindly download the 2nd word document
smunga | Sep 21, 2021, 16:40 PM
Hi, Oh great will have a look now :)
User_39030 | Sep 21, 2021, 17:44 PM
Hello, any chance you have some time to go through it?
User_39030 | Sep 21, 2021, 17:46 PM
Hi dear kindly go through it .On my end i have explained and am certain the final grade is an A.You can ask me any question.But i mAY TAKE SOME TIME TO reply am HANDLING ANOTHER MED EXAM CURENTRY.sORRY FOR THE TYPO MULTITASKING
smunga | Sep 21, 2021, 17:55 PM
Hello dear have you gone through the paper?
smunga | Sep 21, 2021, 18:03 PM
Hi, sorry the messages take some time to go through, I was hoping you could tell me how questions 4 & 7 came about, I also wanted to know how to do the calculations for it. the flow diagram for question 6 and the rest I think I can understand, Sure it's fine, you can take your time replying, no rush...
User_39030 | Sep 21, 2021, 18:14 PM
Question 2 as well, its just the calculations id like to know, would help me in learning and understanding the paper also, if you want, in red you could type up how you came about the calculations in the same document next to the questions, I could read through them and also do it myself as well, Would help me understand whats going on to !
User_39030 | Sep 21, 2021, 18:16 PM
ihave completed the exam.let me answer your questions dear
smunga | Sep 21, 2021, 18:35 PM
n question 6 the first row represents the patient 1 serum dilutions The second row is the duplicate of patient two serum dilutions The third row are the duplicate of negative control serum dilutions The 4th row is the virus only ✅so as you increase the dilutions in each well the number of plaques identified reduces
smunga | Sep 21, 2021, 18:35 PM
in question 2 based on the excel results presented in the Elisa test they were triplicates of each sample .So 1,2,3 represents first sample .4,5,6 represents sample 2 .7,8,9 represents positive control and 10,11,12 represents negative control .In each dilution e.g A you add the each three results and find the average by dividing with three where you come up with the answer .It's repeated in all dilutions to H
smunga | Sep 21, 2021, 18:36 PM
continuation q2(After that you copy the results to excel where you come up with the graph)
smunga | Sep 21, 2021, 18:38 PM
in question 4 you pick any absorbance value in the graph and it must be 0.1 greater than negative control e.g 0.40 Then find dilution of the solution Dilution =1:100iu what about 46.25 is you find it to 0.4625. Then calculate the reciprocal of it to find the titre which is 2.162
smunga | Sep 21, 2021, 18:43 PM
continuation q 4 You'll find out that when you also reciprocate the absorbance value its 2.5 which is between titre range of 2 and 3
smunga | Sep 21, 2021, 18:46 PM
In question 7 count the plaques in sample 1 dilution 1/40before intubation ....its 15 .After intubation it comes to 7 ... calculate the percentage reduction which is 53.3% Also in sample 2 at a dilution of 1/40 before intubation you'll find out that only 10 are visible.After intubation 5 are only visible ..the percentage of reduction will be 50%.. this gives reciprocal of serum dilution
smunga | Sep 21, 2021, 18:46 PM
Okay so for question 6, it says to document the "mean" absorbance? so how do i do that to get those numbers that are in the table?
User_39030 | Sep 21, 2021, 18:51 PM
Oh right sorry i meant question 2, i just understood that
User_39030 | Sep 21, 2021, 18:52 PM
for question 4, how do we calculate the reciprocal?
User_39030 | Sep 21, 2021, 18:53 PM
for question 4, how do we calculate the reciprocal?(its 1/number )eg the reciprocal of 10 is 1/10 which is 0.1
smunga | Sep 21, 2021, 18:57 PM
Dear i also have a favor to ask you would you kindly award me an all star rating and leave a comment with human anatomy it would really mean a lot to me EG(Smunga great with HUMAN ANNATOMY O R Great with Biomedical Science. Kindly
smunga | Sep 21, 2021, 19:00 PM
are you still there friend ?Hope i have assisted you on the areas you had some slight difficulty .Dont forget to rate the task all star.
smunga | Sep 21, 2021, 19:50 PM
Hi, im here, i just have a few more questions i want to clear, how did you normalise the results?
User_39030 | Sep 22, 2021, 19:42 PM
Also for the reciprocal did you mean 1/0.4625 ?
User_39030 | Sep 22, 2021, 19:44 PM
i think we needed to normalise the results as well, but how did you do that ?
User_39030 | Sep 22, 2021, 19:46 PM
Normalise results for question 2 i mean
User_39030 | Sep 22, 2021, 19:48 PM
Let me type a detailed response for each of the questions you have raised.
smunga | Sep 22, 2021, 20:00 PM
If you want to normalize the 'titre'" you have to take a control pool serum (that should from all category) in all plate and convert OD in to ELISA unit. If you want to declare positive or negative from the OD value you have select a cut off value negative.Thats why the point chosen should be 0.1 greater than the negative test
smunga | Sep 22, 2021, 20:04 PM
Yes please! could you please make a detailed response for question 7 as well Sorry and thank you again :D
User_39030 | Sep 22, 2021, 20:06 PM
so does that mean the results were already normalised? or how does that work when i have to show calculation?
User_39030 | Sep 22, 2021, 20:07 PM
Yeah the reciprocal is 1/0.4625
smunga | Sep 22, 2021, 20:12 PM
Q}so does that mean the results were already normalised? or how does that work when i have to show calculation? ANSWER}Yeah when the average was done that means it's already normalised
smunga | Sep 22, 2021, 20:13 PM
I have a questions yesterday i requested you release the funds you were offline .Will you release the funds today?
smunga | Sep 22, 2021, 20:15 PM
are you there?
smunga | Sep 22, 2021, 20:17 PM
Just was hoping to get a clearer understanding of how the calculations came about with working out, Yes! I was going to release the funds today since its due date today anyway,
User_39030 | Sep 22, 2021, 20:17 PM
Also happy to give you your 5 star and review as you're also helping me understand as well :D
User_39030 | Sep 22, 2021, 20:20 PM
do you have an6y yother querry ?I want you to still assist you in all ways possible.But rest assured the task is okay and i belive it an A grade .I gave it my all
smunga | Sep 22, 2021, 20:21 PM
So if you could give me a detailed response on Q2 - so how you normalised the results, like showing the working out? you don't need to do for each result but just one or two so i can see how it came about, that way i also have an understanding. Q4 - Q5 - how were the percentages calculated as well Q7 - i also want to know how to work the solution out here as well and thats it :)
User_39030 | Sep 22, 2021, 20:26 PM
As for question 2 column 1,2,3 averages give the Normalised sample 1..... column 4,5,6 average gives normalised sample 2 averages of column............7, 8,9, gives the Normalised positive control and average of column 10,11,12 gives negative control .For example to find normalised results for dilution A in sample 1 you add 0.071+1.203+1.198 =2.472 it's qverage is done by dividing the answer by 3 =0.824 which is the normalised results
smunga | Sep 22, 2021, 20:53 PM
For question 5 what was basically meant is the sensitivity to N and S protein antigen .The sensitivity of Elisa test to N protein needs it to be more chronic that means the sample should be collected 20 days from the day the first sign was observed. For our case the sample was collected 10 days after the first symptom that means its sensitivity to N protein is low
smunga | Sep 22, 2021, 20:55 PM
Ohhhhhhh
User_39030 | Sep 22, 2021, 21:01 PM
have i simplified it dear ?let me answer the other question
smunga | Sep 22, 2021, 21:03 PM
In question 7 count the plaques in sample 1 dilution 1/40before intubation ....its 15 .After intubation it comes to 7 ...calculate the percentage reduction which is 53.3% 100% of plaques=15 how about 8 cells its 53% Also in sample 2 at a dilution of 1/40 before intubation you'll find out that only 10 are visible.After intubation 5 are only visible ..the percentage of reduction will be 50%.....if 100%of plaques count is 10 how about 5 it gives 50% gives reciprocal of serum dilution
smunga | Sep 22, 2021, 21:09 PM
i guess now all is well.I request you award me an all star rating as promised and leave a positive comment
smunga | Sep 22, 2021, 21:11 PM
are you there?
smunga | Sep 22, 2021, 21:12 PM
Yes I'm here, il release payment now and give you your rating, if there are any questions can i come back to you?
User_39030 | Sep 22, 2021, 21:48 PM
sure and in furure task .Kindly contact me i am ready to assist
smunga | Sep 22, 2021, 21:52 PM
Yes! Thank you so much for everything :)
User_39030 | Sep 22, 2021, 22:18 PM
Thanks to dear .Your feedback is very important would you kindly award an all star rating and leave a comment thanks alot
smunga | Sep 22, 2021, 22:26 PM
If its okay id like to do that tonight :)
User_39030 | Sep 22, 2021, 23:17 PM
Oh for question 7, how do we calculate the percentage reduction as well!
User_39030 | Sep 22, 2021, 23:19 PM
Hey Smunga, how did you end up ding the log dilution?
User_39030 | Sep 23, 2021, 15:53 PM
Hi dear am abit far from my pc.But i will be home in 30 min(out shopping )But i willrespond to the question.In the meantime would you rate the task as promised yesterday?
smunga | Sep 23, 2021, 16:15 PM
Ya no worries !
User_39030 | Sep 23, 2021, 16:26 PM
I just left feedback :)
User_39030 | Sep 23, 2021, 16:32 PM
A log dilution is a tenfold dilution, meaning the concentration is decreased by a multiple of ten. To complete a tenfold dilution, the ratio must be 1:10. The 1 represents the amount of sample added. The 10 represents the total size of the final sample.so you hsve to find half log inverse of the chosen point in the graph o.40 which is 2.51 which is within normalised trange of 2-3
smunga | Sep 23, 2021, 17:00 PM
is this where it says "empty wells" etc
User_39030 | Sep 23, 2021, 17:18 PM
There were no empty Wells for our case....incase of empty Wells we'll do half log
smunga | Sep 23, 2021, 17:56 PM
So in page 7 of the practical 6 pdf INDIRECT ELISA, it shows empty wells?
User_39030 | Sep 23, 2021, 18:59 PM
oh no i think you are right
User_39030 | Sep 23, 2021, 19:01 PM
Hey Smunga, so if you go to page 13 in the week 6 practical PowerPoint, there is also another set of results, and it says empty well etc ?
User_39030 | Sep 23, 2021, 23:47 PM
are you able to explain question 7 to me again please, I'm still a little confused about it
User_39030 | Sep 24, 2021, 01:36 AM
step 1 In question 7 count the plaques in sample 1 dilution 1/40before intubation its 15 (end of step 1) (STEP 2).After intubation it comes to 7 ... calculate the percentage reduction which is 53.3% (END OF STEP 2) (STEP 3) Also in sample 2 at a dilution of 1/40 before intubation you'll find out that only 10 are visible. After intubation 5 are only visible ..the percentage of reduction will be 50%.. this gives reciprocal of serum dilution(I DONT KNOW HOW EELSE TO EXPLAIN THIS )
smunga | Sep 24, 2021, 03:54 AM
I understand it now Thanks. Also, I noticed there weren't any x and y-axis titles for the graph? I've also got given a few notes in regards to the assessment, il send it here so you can see.
User_39030 | Sep 27, 2021, 03:32 AM
Plaque Assay I did not give you information on how much we diluted the virus to begin with but all the wells got the same amount of virus so work from the raw values (plaques) you count. Percentage reduction = ((average virus only – sample) / average virus only) x100 Titre- reciprocal of the virus that gives you 50% neutralisation. If neutralisation is higher than 50% at the highest dilution of the serum used report that the titre is higher than the reciprocal of the highest dilution used.
User_39030 | Sep 27, 2021, 03:35 AM
Indirect Elisa: Blanks- we have a row of empty wells, that got assay reagents but not abs- Row A that we designated – please use that row. Average the readings and subtract the average from your wells. You have several controls in the assay in row 6-7 the positive serum sample and row 8 two different controls we included to see if the assay was specific. As mentioned previously rows 9-12 are not used, the readings are the absorbance value read in the empty plastic wells. You don’t need to bother with these wells at all.
User_39030 | Sep 27, 2021, 03:36 AM
You were given three options to determine your values here Take 2- or 3-fold the average background levels as the cut-off and report anything above this as positive below is negative Calculate the mean value + 3-SD for the negative wells and take this as the cut-off. Use same dilution in all samples that best differentiates +ve and –ve samples What is important is to use a uniform criteria during a given study and report this criteria in your results The criteria you chose will influence if you think sample A and B are both positive
User_39030 | Sep 27, 2021, 03:36 AM
Are you able to send me the graph with the x and y axis titles please :)
User_39030 | Sep 27, 2021, 03:37 AM
Right so for question 7. how did you get the numbers of 15 and 7 is what i mean
User_39030 | Sep 27, 2021, 03:48 AM
OH LOL, im so confused i saw the plaqe in the result
User_39030 | Sep 27, 2021, 04:06 AM
Sorry for all the messages but I think the ELISA is testing for anti-corona virus antibodies in the patient and the plaque assay tests for the patient's ability to neutralise covid using their antibodies?
User_39030 | Sep 27, 2021, 05:51 AM
wouldnt that mean patient 1 was positive? as the results were similar to the positive control result
User_39030 | Sep 27, 2021, 05:56 AM
Sorry Smuga, can we organise a time where you are free, some things are a little bit unclear to me, was hoping to sort it out, let me know if you're okay with this
User_39030 | Sep 27, 2021, 05:59 AM
hey, sure. am taking a students exam , but i will be done in the next two and half hours.
smunga | Sep 27, 2021, 06:04 AM
Thank you very much !
User_39030 | Sep 27, 2021, 06:13 AM
There is alot of information you have uploaded what is your question?
smunga | Sep 27, 2021, 07:02 AM
So for the results you state that there were no antibody detected? but im seeing that patient one was positive for anti-corona antibodies and patient two has the presence of anti coronavirus antibodies but at less than half the amount of patient one? I didn't really understand the results to be honest, maybe you can explain it to me ?
User_39030 | Sep 27, 2021, 08:26 AM
Hey smuga Mm happy to make another assignment assigned to you with payment as well, considering you are familiar with this, id like to go over the same report
User_39030 | Sep 28, 2021, 02:42 AM
Hello am sorting out some small issue but i am ready to assist you on the task Once the issue is resolved .Thank you for choosing my services again. As always expect top quality assignment delivered on time. Am also available for follow up question .Kindly inform me when you post the job. Thank you
smunga | Sep 28, 2021, 02:50 AM
hi Smuga are you familiar with the subject histology ?
User_39030 | Oct 01, 2021, 03:24 AM
tissue staining etc?
User_39030 | Oct 01, 2021, 03:24 AM
yes sure did it in my undergrad med class.You have a task ?
smunga | Oct 01, 2021, 03:30 AM
Yes Smuga, i do have a task. But was wondering if you would be able to complete it in under like 6-7 hours? there are about 4 parts, but I was hoping if you could do part 1 and 2. Part 1 asks What is the Composition and description of your tissue element stained by Harris’ H&E. and part 2 asks Using the best-specialised staining technique for carbohydrates and mucosubstances of your tissue explain the chemistry of the interaction of the dyes/labels with your tissue elements. The tissue i have is stromal extracellular matrix. What do you think? If you think part 3 can be done as well, please feel free to do so, i am willing to increase the amount Let me know whatt you think asap
User_39030 | Oct 03, 2021, 01:00 AM
hello friend sorry for the late reply am willing to assist you all through .Kindly post the task and i will bid on it thank you
smunga | Oct 03, 2021, 01:32 AM
Sure ! will post in 10 minutes if its okay :)
User_39030 | Oct 03, 2021, 01:45 AM
Hello friend are you there? Have you posted the task?.
smunga | Oct 03, 2021, 02:09 AM
Yes, please have a look
User_39030 | Oct 03, 2021, 02:16 AM
i can see the task on the site ?WHAT IS THE TITLE OF THE JOB?
smunga | Oct 03, 2021, 02:22 AM
Anatomical PATHOLOGY questions on tissue staining
User_39030 | Oct 03, 2021, 02:24 AM
Did it show up ?
User_39030 | Oct 03, 2021, 02:25 AM
yES and have placed my bid
smunga | Oct 03, 2021, 02:27 AM
Kindly assign me the task.I start
smunga | Oct 03, 2021, 02:31 AM
How many hours remaining ?( i will do both part 1 and 2)
smunga | Oct 03, 2021, 02:31 AM
Hey smuga, is it ok if you can lower the bid for now, just so my card can go through and I can assign you the assignment, then I'm going to add the extra bonus to add up to 90$ in for you once that's done :) Hope this is ok
User_39030 | Oct 03, 2021, 02:33 AM
It is due before midnight tonight, and it's 1.30pm afternoon here,
User_39030 | Oct 03, 2021, 02:38 AM
Part 1 and two is great :)
User_39030 | Oct 03, 2021, 02:38 AM
Kindly assign me the task to Quality assignment
smunga | Oct 03, 2021, 02:41 AM
Hey Smuga, I want to assign the task although I needed the bid to be lower please, once i assign task il add in the rest of the funds straight away.
User_39030 | Oct 03, 2021, 02:43 AM
Assigned task! Sorry about the funds, i have to keep adding in bit by bit :'(, please bear with me in the meantime
User_39030 | Oct 03, 2021, 02:52 AM
Its okay already working on the task .As explained my girl friend is abit too strict so kindly don't forget to rate her ALL STAR .I had to make a Special request since my account could not allow me change the amount. Also promised her you would leave an amazing comment like you did last time.
smunga | Oct 03, 2021, 02:59 AM
No worries! i can do all that for you :) Thank you again, you've been amazin
User_39030 | Oct 03, 2021, 03:01 AM
Hello
smunga | Oct 03, 2021, 06:21 AM

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