Added Responsibility - Week 2

Finally, a little more responsibility.  The end of my first week in lab finished with a bang as I was invited to take the validation of a new assay to completion.  Because this lab processes active patient samples, each test and modality used needs to be validated before hand.  This involves running the new and old test side by side and comparing results, specificity, and sensitivity.  Because all samples used in a validation are from previous patients, not current ones, I was able to run some of the tests.  The new assay we were testing was for HepB, both the initial antibody testing and the confirmatory assay.  In total we ran about 500 samples (at least 3 times each) and ended up with a 14 page excel sheet to prove it.  This project was a major improvement over the beginning of my week and offered some hope for the future.

My expectations were met toward the beginning of my second week.  Having observed most of the benches and tests in the lab, my PI Dr. Hodinka decided that I should get more hands on experience.  One of the tests he wanted me to run was a Western Blot HIV confirmatory assay.  This test detects proteins in a sample (HIV antibody in this case) by binding them to a strip.  Each band on the strip represents a different protein and the intensity of each band can indicate the severity of the virus present.  The blot that I ran compared two lots of controls; a high positive, low positive, and negative control.  Because this test is slightly outdated, it is preformed over night and is very labor intensive.  I had to mix my reagents and manually add them to my plate.  

HIV Western Blot      

The most exciting part of the week in my opinion was the indication that my project would begin sometime next week.  Because most advances made in the field of clinical virology in the future will be in the molecular testing area, Dr. Hodinka will most likely give me a project involving PCR of some kind.  After months of having no clue what I would be doing in the lab, this little inkling has me pretty excited.  Being the "gold standard" of molecular testing, RT-PCR (real time polymerase chain reaction) is highly specific and is a relatively efficient way of identifying and diagnosing viruses present in sample.  Differently from traditional end point PCR, real time PCR can detect amplification as it is happening and accurately report how severe an infection is based on the number of cycles it took a positive sample to pass a threshold.  This is a very useful piece of information for clinicians when making a diagnosis.

Overall I am very excited about the remainder of my time in the lab.  The people who work here are very friendly and always willing to explain something or offer insight when I need it.