ELISA Test for Virus Detection

enzyme-linked-immunosorbent serologic assay Test for virus DetectionZohrah MiakheylImmunology practicalIntroductionViruses ar fine living organisms that requires living host for its selection and pitiablelife lead to transmission establishment if it fully invades the host cells taking over the cell machinery to produce virus. on that point are many different viral contagious distempers including measles, epidemic parotitis, rubella, and cytomegalovirus. measles is an infection of the respiratory system campaignd by the morbilli virus. The measles virus is very transmittable and stub easily spread via sneezing, coughing and personal contact. rubeola is cognise by the red-brownish rash starting on the face and spreading to the expect of the dust. Other symptoms of measles virus include fever, diarrhoea, runny nose, conjunctivitis and ironical cough (Permar et al, 2006). Mumps is an infection fixd by the paramyxovirus. This viral infection is too highly contagi ous like morbilli and spreads in addition via sneezing, coughing and bear contact. Mumps usually causes swollen salivary glands, fever, dry m discloseh and difficulties in swallowing. rubella is an infection caused by rubella virus, member of the genus Rubivirus. The spread of the virus is similar to measles and Mumps. Most common symptoms of this infection are a pink, light red rash lasting up to iii days, diffuse fever, cold cough, sore throat, conjunctivitis and joint pain in adults (Stock, 2012). In contrast to Measles, Mumps and Rubella, Cytomegalovirus (CMV) usually dont own any symptoms and hence most people being septic may non know that they have CMV. People who are being infected often develop soft symptoms similar to flu or glandular fever. This viral infection tail commence serious in babies infected before birth, because this virus can be transmitted from mother to foetus. Cytomegalovirus is a virus belonging to herpes family of viruses. This virus is spr ead via body unsounds (saliva and urine) and personal contact (Griffiths, 2002). The above stated viral infections usually arrive at childhood, but it can occur at any time. Measles, Mumps and Rubella are often protected by giving the children MMR inoculation at 13 months and a booster is often given to three to five age old children. To find out from what the four perseverings are mislaying from ELISA technique was applied to detect antiviral agent antibodies in endurings serum for the viral infections Measles, Mumps, Rubella and Cytomegalovirus.MethodResults elaborate 1 microtitre plate showing the moments of the four affected roles tested.The ELISA technique shows that persevering 1 is incontrovertible for Measles (IgM) and CMV (immunoglobulin G), patient 2 is confident(p) for Measles and Rubella (immunoglobulin G), diligent 3 is col later(prenominal)al for measles (IgG) and patient 4 is exacting for CMV (IgM) and Measles, Mumps and Rubella (IgG).Discussion quadr uple patients were tested for viral infections using ELISA technique and the results showed absolute for several(prenominal) patients for some of the viral infections. The ELISA results obtained were good and no errors were make. This might be due to accurate practical skills such as pipetting, appropriate selection of the wells, proper emptying and washing of the plate. Furthermore accurate serum dilution and meter has also added to the accuracy of the results. Controlling the experimental conditions such as brooding time has also lead to improved results. To obtain better results saturation measurement, dilutions, experimental conditions can be improved further. The volume measurement can be improved by accurately use of Gilson pipettes by ensuring there is no air bubbles and the pipettes are fully calibrated. Experimental condition can be improved by carefully monitoring the incubation duration.Patients case studyPatient 1A 56 age old patient was admitted with dense reddish brown maculopapular rash, headache, low grade fever, spot rash, mild group AB discomfort and diarrhoea, high temp, mild conjunctivas symptoms. Throat swab done by the GP didnt let loose any severe symptoms, but was referred for antiviral antibodies screening examination. aft(prenominal) playing antiviral antibody screening the results showed positive for measles (IgM) and CMV (IgG). IgM is made during immune reception in the too soon demo of infection and is principally form in lymph fluid and slant where it situations as neutralizer against harmful agents. Whereas IgG is made in the late stage of infection and cheques in the body for a long period of time. Due to its long survival period in the body IgG is useful for passive immunization. IgG is mainly found in blood, lymph fluid, cerebrospinal fluid and peritoneal fluid and plays a vital role in humoral immune response by activating the complement system (Lima, 2012). The sign and symptoms of the patients is similar t o measles therefore indicating that the patients might be suffering from measles and might be in the early stages of the infection due to IgM found in the serum sample and macolopapular rash is also the sign of the infection at early stages. The mild abdominal discomfort, diarrhoea, and cold-like symptoms also indicates that the patient might suffer from CMV infections, but since IgG is found in patients serum sample, the infection might also be the result of immunisation and prior infection. All the symptoms of this patient are also the symptoms of measles therefore this patient might suffer from measles in the early stage of it, because of the antibody IgM and the reddish macolopapular rash is also found in the early stage of the infection) . Currently there are no means of preventing the positive measles infection, but the symptoms can be reduced using pass medications such fever-reducers, antibiotics, and vitamin A supplements (Permar et al, 2006).Patient 2A healthy 8 years old boy was examined by GP in a routine check and was referred for full blood look ats. From the physical examination, the GP found that the boy was having faint cracking-like sound due to sever cold that he has been suffering since 3 weeks. Eczema-like rash was also observed, which was treatable by E45 creams, but the cause of this symptom was not clearly known as he has no family history of eczema. After his full blood deliberate revealed no ab commonplaceities and the cause of eczema was not known, his GP referred him for antiviral antibodies screening. The screening result showed that the boy was positive for Measles and Rubella (IgG) infections. The signs and symptoms of this patient appears to be different from the sign and symptoms and Rubella and Measles apart from the cold that the patient was suffering from. One of the premier signs of Measles and Rubella infection is red-pinkish rash, where in Measles it appear to be observable for a week and in Rubella it lost for at least(prenominal) 3-7 days, but varies from patient to patient. The patient always had eczema-like rash, which means that the patient might not be infected by Measles or Rubella. The positive results obtained from antiviral screen might be due to immunisation and prior infections. Since the patient is not suffering from virus infection, the doctor ca refer him to Dermatologist for eczema examination. Since he is suffering from cold, the patient can orchestrate cold reducing medications (Gen,1999).Patient 3A 26 years old healthy female attend her GP for confirmation of her gestation period test after she tested at piazza using home pregnancy test about a week ago and got positive results. She believes she is about 28 days expectant, but this is her fist confirmation although she has been significant in the past and had early stillbirth about 10 months ago. GP preformed second pregnancy test and the positive results confirmed that she is pregnant. Since she was only 4 years old when moved to UK from Tajikistan, her medical record is available from the age of 6. Due to incomplete medical records, the patient is advised to carry out blood test so that her vaccination status can be known, therefore antiviral antibody screen was carried out and the results appear positive for Measles infection. The development of MMR vaccination appear to be very successful against this viral infection and due to improve and advancement of MMR vaccination, Measles infection has become less common now, but it shows to be dangerous among pregnant women if occur. This is because Measles might lead to miscarriage, stillbirth or early delivery if the pregnant individual is not immune against the infection. Although patient 3 do not peril any sign and symptoms, but the antiviral antibody screen shows that she is positive for this infection. This positive result suggests that the miscarriage that she had might be due to Measles infection. However this cannot be said with certainty , because of her incomplete medical history. The antiviral positive result might be due to early infection or immunization. Since her medical history is not fully known, she probably needs MMR vaccination to make sure she is immunized against the viral infectionNormally the signs and symptoms of Measles infection tend to appear between 6 and 21 days. . and so its important that she is aware of any signs symptoms such as fever, cold and any skin rash that might appear at later stages of Measles infection. If she is infected then human normal immunoglobin (HNIG) treatment can be carried out to reduce the symptoms of infection. However this does not mean that miscarriage can be prevented (Permar, 2006).Patient 4A 43 years old male who had kidney transplantation 9 months ago due to kidney failure, which was caused as a result of Type 1 Diabetes Mellitus, which was diagnosed at the age of 11. The patient had a good diet and controlled blood glucose, but due transport in life style and alcohol consumption and irregular insulin institution his blood glucose level has changed and become unpredictable. This life style has led to impaired kidney functioning, which resulted in estimated Glomerular Filtration Rate (eGFR) of 30ml/min. After monitoring eGFR about 3 months, a steady reduction was observed until the age of 39 with eGFR of 10ml/min. Although the patient was under dialysis, he was introduced to ambulatory peritoneal dialysis (CAPD) while waiting for capable kidney donor for transplantation. After a few months CAPD treatment, he was referred to haemodialysis, which was carried out 3 times a week, because he could not get on with CAPD. After successful single kidney transplant, the patient shows to recover well from the surgery and the kidney appeared to function well. Following the checkups, the patient appeared to feel better and showed good recovery. However, the patient complained of broadly speaking unwell feelings with severe cold/ flu, where he has s uffer for at least couple of weeks. Chest pain, abdominal cramps with diarrhea was also reported on this visit and physical examination shows some abdominal tenderness with no swelling or any other tenderness and normal blood pressure. The Auscultation examination however showed rattle and wheezing sounds. Microbial culturing was performed on stack and mid-stream urine samples a land with full blood count, creatinine measurement and an antiviral antibody screen examination.The microbial culture from urine samples revealed nothing significant and eGFR shows to be similar to last check up (10ml/min). The full blood count however shows that the Haemoglobin concentration was 12g/do, which is lower than the normal digress (normal range 13-18g/dL) and this is thought to be as a result of impaired glomerular filtration rate, which is caused due to kidney failure and his haemoglobin concentration might be also low due to his kidney-transplantation and the immunosuppressive medication tha t he receives which causes mild bone-marrow crushing causing low hemoglobin concentration (Joist et al, 2006) . All other tilt however appear to be within the normal ranges indicating nothing significant. exclusively the white blood count appears to be slightly higher than the normal range. The normal white blood count is 4-1110 to the 9/L, whereas the patients count is 11.110 to the 9/L. This indicates that the patient might be suffering from viral infection. This is intelligible from the antiviral antibody screen examination, where the patient appeared to be positive for CMV (IgM), Measles, Mumps, and Rubella (IgG). Therefore it is the Measles infection that causes the while blood cell counts to alter. In general viral infections disrupts the bone marrow function temporarily and this leads to transformation in white blood cell counts. The severe cold/flu, abdominal discomfort and diarrhea that the patient is suffering from is also the signs of CMV infection, this is because i ndividuals who afford organ transplantation are more susceptible to CMV infection (Rafailidis et al, 2012).The reason that the individuals are susceptible is because the medication they received reduces the immune system response to avoid rejection of kidney transplant (Cordero et al, 2012). However the patient does not display any symptoms for Rubella, Measles and Mumps, but the positive antiviral screening result might be due to previous infection or immunization. His crackling and wheezing sound might be result of cardiovascular disease that he might suffer from because of his lifestyle Kataoka, 2008).In order to stay healthy the patient needs to improve his lifestyle by improve his diet by reducing/stopping alcohol consumption. If the symptoms of CMV becomes worsened than antiviral drug can be used. If the immune system is jury-rigged weakened then there is no need for any antiviral drug, because the patient will recover from the infection when the immune system is recovered. harken of referencesCordero, E. et al. (2012). Cytomegalovirus disease in kidney transplant recipients incidence, clinical profile, and risk factors. Elsevier. Online 44 (3) p. 694-700. Available from http//www.ncbi.nlm.nih.gov/pubmed/22483471. Accessed 15/02/2015Gen. (1999). Immunoglobulin (IgG) and (IgM) Antibody Responses to Rabies Vaccine .Lister Institute of load Medicine, Elstree, Hertfordshire, U.K. 40 (10), p595-604.Griffiths, P. D . (2002). Cause and Treatment Of Cytomegalovirus Infection.Journal of Antimicrobial Therapy. 49 (2), 243-255Joist, H. Brennan, D.C. Coyne, D.W. (2006). Anaemia in the kidney-transplant patient. Journal of The American rescript of Nephrology. Online 13(1) p. 4-10. Available from http//www.ncbi.nlm.nih.gov/pubmed/16412965. Accessed 11/03/2015.Kataoka, H. Matsuno, O. (2008). Age-Related Pulmonary Crackles (Rales) in Asymptomatic Cardiovascular Patients. Annals of Family Medicine. Online 6(3) p.239-245. Available from http//www.ncbi.nlm.nih.gov/p mc/articles/PMC2384982/. Accessed 15/03/2015Lima, R. C. et al (2012). Interpretation of the presence of IgM and IgG antibodies in a rapid test for dengue analysis of dengue antibody prevalence in Fortaleza City in the 20th year of the epidemic. Journal of the Brazilian Society of Tropical Medicine. Online. 45(2) p. 163-167. Available from http//www.ncbi.nlm.nih.gov/pubmed/22534985. Accessed 22/03/2015.Permar, S. R, Griffin, D. E and Letvin, N. L. (2006). 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