...Serial Dilutions & Antibody Titers CLS 240 May 21, 2014 LABORATORY EXERCISE – WEEK 2 Serial Dilutions Principle A set of dilutions where the dilution factor is exactly the same at each step, is known as a serial dilution. Serial dilutions are commonly used in the clinical laboratory to determine the strength or titer of a particular antibody in patient serum. This information can be used to help diagnose the disease state of the patient, which will help the provider develop an effective treatment plan. Serial dilutions are created with a known quantity of both serum and diluent. When making serial dilutions it is import to ensure the zone of equivalence is maintained. The zone of equivalence is where there are an equal amount of antigens and antibodies within the dilution. If the zone of equivalence is not obtained and there are too many antibodies in the dilution it is known as prozone, whereas too antigens it is known as postzone. Hemagglutination studies use the concentration of antibodies to help diagnose patients. When a patient first becomes ill his/her titer will be low, which is known as the acute phase. In the acute phase the titer is low because the patient has recently become ill and the patient’s body has not had enough time to develop antibodies. It is estimated within 2 weeks the patient will enter the convalescent phase where the titer will become increased, suggesting the...
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...asymptomatic[2]. Chronic gastritis is also a known risk factor for adenocarcinoma, but it is thought that is takes many years of chronic infection before cancer cells develop[3]. H.pylori is also believed to play a role in other GI conditions such as gastroesophageal reflux disease and gastric lymphomas [3]. H.pylori can be diagnosed through invasive procedures such as endoscopy with a biopsy[3], or with non invasive procedures that include fecal analysis, urea breath test, and serology[2]. Endoscopy provides a large amount of information for diagnosis in regards to which antimicrobials the bacteria is susceptible to, but it is the most invasive diagnostic procedure for the patient, and is usually unnecessary, with less invasive procedures proving to be just as adequate for diagnosis of H.pylori. A examination of a serology specimen may actually be more indicative of the degree of infection. Since the serology test takes a sample from the entire stomach, as opposed to the smaller sample size of a biopsy, a serology sample test is actually more sensitive and can more easily detect the presence of H.pylori [3]. Another effective diagnostic test utilizes H.pylori’s urease virulence factor. A urea breath testis done when a patient orally ingests urea that is tagged chemically, usually with a carbon isotope, if H.pylori is present in the stomach, it’s urease will convert the urea(CH4N2O) to ammonia(NH3) and carbon dioxide(CO2)[3][4]. The carbon dioxide(CO2) will have the carbon isotope tag...
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...(i.e. a severe and extreme allergic reaction). Other potential cyst complications include fistulas leading to bronchial or biliary obstruction, embolism of cyst content and bacterial superinfections. Diagnostic approaches Due to its availability and convenience, the introduction of ultrasound in the clinical practice in the late 1970s has substantially improved the detection of cysts in the liver and abdomen. World Health Organization developed a standardized classification system (pervasive in modern medicine) which divides hydatid cysts into six stages: CE1 and CE2 (active stage), CE3a and CE3b (transitional stage), and CE4 and CE5 (inactive stage). Serology is also helpful, but its usefulness is limited by a lack of sensitivity and specificity. Still, the combination of ultrasound technique and confirmatory serology represents a standard approach for epidemiological and clinical surveys. In unclear cases, diagnostic puncture of the cyst is performed by an experienced examiner. Treatment and prevention In general, four different modalities are available in the treatment of hydatid disease: antihelminthic treatment, percutaneous drainage techniques, surgery and “watch and wait” approach. However, the management of patients depends not only on the individual case, but also on local expertise and available resources. Surgery is indicated when there are large hepatic cysts (with multiple daughter cysts) or superficial hepatic cysts with the potential of spontaneous rupture...
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...Running Head: Blood Splatter Analysis Blood Spatter Analysis Kenisha Callahan AJS 592 Dr. Steven Gilbert Blood Spatter Analysis Blood spatter analysis is “the systematic assessment of the visual patterns of bloodstains at crime scenes based on the physics of fluids” (Brodbeck, 2012, p. 51). It has been studied since 1895 when Dr. Eduard Piotrowski published "Concerning the Origin, Shape, Direction and Distribution of the Bloodstains Following Head Wounds Caused by Blows." The next acknowledgment of BPA was in 1955 when Dr. Paul Kirk entered blood splatter evidence in the State of Ohio v. Samuel Sheppard case. Dr. Herbert MacDonell started training personnel of law enforcement about blood spatter analysis in 1971. Since that time, bloodstain pattern analysis has grown and continues to develop. Blood pattern analysis is used in a lot of crime scene investigations and can hold valuable information. Blood spatters can indicate important clues such as, the number of blows to the victim, the force of the blows, weapon used, and position and/or movement of the victim and suspect. Blood spatter can be classified into three categories, low velocity, medium velocity, and high velocity (Saferstein, 2011). Low velocity spatter consists of large separate or compound drops with diameters measuring three millimeters or more. It is usually the result of dripping blood. Medium...
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... Zoonotic pathogens : - B.Abortus / B.Melitensis Symptoms: -long term fever - Undulant fever - Complication in bone manifestation: bone pain ; arthritis - Re-current infection is common - Virus like infection Identification: - Oxidase + Urease+ -Serology test IgG/ IgM -gram negative rods,Very small cannot be seen under microscope -Slow grow better isolate with CO2 and increase humidity. -Sampled from blood (5 to 7 days b4 giving a negative report) -cultured on chocolate agar(enrichment media). - Wright test (like widal “ anamnastic reaction” ) Transmitted via contaminated food Milk products not from human to human : can be killed at high temperature: - tantalization : 50º - Pasteurization: 70 º - Sterilization : 100 º Treatment: - Tetracyclin - Doxycyclin - Rifampin - For 6 weeks and more - Vaccine is available for animals. Vibrio V. cholerae, V. parahaemolyticus, V. vulnificus Features: - Gram negative - Fermenter bacilli - Facultative anaerobes - Live in halophilic places Identification: - Enrichment medium - alkaline peptone broth Vibrios survive and replicate at high pH - Other organisms are killed or do not multiply - Selective/differential culture medium - TCBS agar - V. cholerae grow as yellow colonies - Biochemical and serological tests Pathogenesis and treatment: Rehydration & supportive therapy Oral Intravenous (IV): - Doxycycline or tetracycline (Test resistance may be developing) of secondary...
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...Close window ------------------------------------------------- Infectious causes of abortion | Infectious factor Common names | Abortion rate | Abortion timing | Recurrence of abortion | Foetal lesions | Samples | Bacterial | Brucella abortusBrucellosisBang’s diseaseZoonosis | Up to 80% of unvaccinated animals infected in 1st or 2nd trimester | 6-9 monthsAbortion or stillbirth 2 wk to 5 mo after infection | Majority abort only once | Placenta: retained, cotyledons necrotic, red-yellow,; area between thickenedCalf: normal or autolytic with bronchopneumonia | placenta, foetus, or uterine dischargeDiagnosis: maternal serology, IFAT for Abs in placenta, bacteria isolation | Campylobacter fetus venerealisVibriosis | >10% | 5-8 months | Uncommon, convalescent cows resistant to infection | Placenta: mild placentitis, hemorrhagic cotyledons and an edematous intercotyledonary area.Foetus: fresh or autolysed; mild fibrinous pleuritis, peritonitis, bronchopneumonia. | Placenta, foetal abomasal contents, vaginal flushingDiagnosis: microscopic detection, isolation | C fetus fetusC jejuni | Sporadic | 4-9 months | Uncommon, convalescent cows resistant to infection | See above | See above | Leptospira interrogans, serovarsgrippotyphosa, pomona,hardjo, canicola,icterohaemorrhagiaeZoonosis | 5-40% | Last trimesterAbortion 2-5 weeks after infection | Immunity to the serotype causing abortion but sensitive to other types | Placenta: diffuse placentitis with avascular, light...
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...(Active) SERIOUS MENTAL DISORDER Mental Retardation (Mental Deficiency), Insanity, Previous Occurrence of one or more attacks of Isanity, Anti-Social Personality, Mental Defects, Epilepsy, Sexual Deviation, Narcotic Drug Addiction, Chronic Alcoholism PHYSICAL DEFECTS AND DISORDER Class A Class B Physical defects, disease or disability serious in degree or permanent in nature that impairs the ability to earn a living as to make them likely to be a public charge MINOR CONDITIONS Class C MEDICAL RECORD 1. Pertinent Medical History: 2. Significant Physical Examination: 3. Chest X-ray report: (for ages 11 years & above) Present recent x-ray film (14x17 inches) 4. Laboratory examination: (attach laboratory reports) a. b. c. d. ( Blood Serology: RPR/VDRL (Ages: 15 yrs. And above) Urinalysis: (Age: 1 yr. and above) Stool (Ova and Parasite) : (Ages: 1 yr. and above) Other examination(s), if necessary ) Not physically and mentally defective or diseased EXAMINING PHYSICIAN / License No.:...
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...Blood type A/B/AB/O Most O+ 39% Least AB- 1% Antigens, usually proteins, are located on the surface of red cells and are responsible for blood-type characteristics. Plasma, which is the fluid portion of blood, is composed principally of water. Rh factor another antigen found on the surface of red blood cells 85% of human population is Rh+ Anibodies are Y-shaped proteins secreted by white blood cells that attach to foreign antigens to destroy them. If you have blood type A, you have Antigens on your red blood cells, and B antibodies in your blood plasma. Those B antibodies will clump if mixed with B antigens. Genotype I(b) I(b) Phenotype Blood type B Heterozygous I(a) I (i) Homozygous I(a) I(a) Chapter 10 Serology is used to describe a broad scope of laboratory tests that use specific antigen and serum antibody reactions The concept of specific antigen-antibody reactions has been applied to immunology techniques for the reaction of drugs of abuse in blood and urine. Forensics of Blood The determination of blood is best made by means of a preliminary color test. A-B-O vs. DNA Seminal stains- sexual contact Blood spatter analyst at crime sense Transfer of blood Arterial spurting----splash of blood---dribble down on the wall Blood drip onto blood---surface Violence of event (energy) Very energetic event-----gun shots Falling drop of blood Spherical------ hit at an angle Multiple blood droplets---plot backwards---point...
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...Summary of activities engaged and outputs in Virology and Molecular Diagnostics Unit, IITA Ibadan in 6 months- Oyekanmi O. Joshua 1. Management and analysis of data generated on expression studies of Yam viruses at IITA using antibody sources: IITA, India (SVU) and Winter (DSMZ). Output: All available data were analyzed and compiled into designated folders. 2. Skype call meeting 'Yam Virus Toolkits Project' Output: Submission of reports of Skype meeting monthly 3. Cloning of Yam virus in Escherichia coli for expression and purification of Yam Virus(YMV&YMMV) CP in Escherichia coli Output: Successful clones were stored at -70oC 4. Cell growth, Expression studies and Purification of Maize Virus(MCMV) CP In Escherichia coli Output: Reproduced recombinant coat protein of MCMV 5. Cloning of Banana Virus(BBTV) CP In Escherichia coli for sequencing Output: Trained PhD student on cloning and successful clones were sent for sequencing. 6. Evaluation of YMV antibody generated Output: Used Direct Antigen Coating ELISA technique to evaluate YMV antibody generated, bleed 5 of India (SVU) source, managed and analyzed the data generated- there was no reaction. 7. Cloning of Yam Virus(Badnavirus) CP In Escherichia coli for sequencing- Output- Trained PhD student and colleague on Cloning and successful clones were sent for sequencing 8. Evaluation of genera Potyvirus antibody to determine the reactions of genera potyvirus antibody against...
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...The natural history of syphilis is very variable. The course of the infection spans many years and may lead to various clinical presentations, which are classified into early (infectious) and late (non-infectious) stages. Early syphilis may be further divided into primary, secondary, and early latent syphilis; late syphilis includes late latent and the various forms of tertiary syphilis. The immune response to syphilis involves production of antibodies to a broad range of antigens, including non-specific antibodies (cardiolipin or lipoidal) antibody and specific treponemal antibodies. The bacterium Treponema pallidum is a spirochetal bacterium that is the cause of syphilis. T. pallidum is a spirochete it can range from 6 to 20 um in length and 18 to 20 um in diameter it requires low amounts of oxygen which it is able to draw from the atmosphere around it. Spirochetes are usually Gram negative but T. pallidum will not show up on a gram stain because the organism is too thin. It can however be detected using special stains such as the Dieterle stain. Spirochetal bacterium have long helical shaped cells the reason for the helical shape if because of the different arrangement of the axial filaments which is known as endocellular flagella the filaments run the length of the cell causing the spiral shape, these flagella are used to move the spirochete. Spirochetes are usually found in liquid environments such as blood, lymph or water. (Encyclopedia Britannica 2010.) Syphilis can present...
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...Daniels Professor Ian Rodway George Mason University Forensic Misconduct: Pamela A Fish Forensic science is defined as the practice of utilizing scientific methodologies to clarify judicial inquiries. The field of forensic science contains a broad range of disciplines and has become a vital aspect of criminal investigations. Some forensic disciplines are laboratory-based; while others are based on an analyst’s interpretation of observable patterns (Kourtsounis, 2009). According to the Innocence project’s website; in greater than fifty percent of wrongful convictions, the use of invalidated or improper forensic techniques played a role in cases; which were later overturned by DNA testing. Unlike other forensic methods, serology has been subjected to rigorous scientific testing to achieve validation. Even still, these methods can be inaccurately conducted or counterfactually conveyed during the trial (Innocence Project, 2012). Since the evidence presented during a trial can be the difference between freedom, and incarceration in many cases and death in some cases, misconduct in the forensic field is not tolerated. Some types of forensic misconduct include exaggeration of statistics, false testimony and laboratory fraud. Evidence offered by forensic scientists is often called “expert testimony”. Nevertheless, what is an expert? Is it someone who has gone to school and earned an undergraduate, graduate or doctoral degree? Alternatively, could it be someone...
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...Background: Mediterranean Spotted Fever (MSF) is characterized by maculopapular rash, fever and eschar at the tick bite site. It is a summer illness caused by Rickettsia conorii that is endemic in the Mediterranean basin. The main known vector is Rhipicephalus sanguineus. Although the majority of cases are benign, complications and mortality can occur. The objectives of this study are to clinically characterize and to delineate predictors of bad outcome in patients hospitalized with Mediterranean Spotted Fever during 23 years. Material/methods: We performed a retrospective cross-sectional study of adult patients hospitalized in an Infectious Diseases Unit between 1992 and 2014. The MSF was defined as positive R. conorii serology, positive R. conorii Polymerase Chain Reaction or clinical diagnosis (typical rash and eschar). The patients were identified by our hospital database (ICD9 082.1) and afterwards clinical records were consulted. The primary outcome of this study was patient outcome (heal or death) and secondary outcomes were Intensive/Intermediate Care Unit (ICU) admission, complications (consciousness impairment, pneumonitis, renal failure, meningitis and gastrointestinal bleeding) and predictors of death. We used the most appropriate measure of central...
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...Forensic science has emerged as a powerful tool in criminal investigations. The success of series such as Crime Scene Investigation (CSI) and Bones has greatly influenced the notoriety of this area of the sciences, mostly including only the physical aspect of crime investigations. This limited view does not demonstrate the diversity contained within forensic science. Internal workings of the bodily functions, chemically and biologically, are just as important in the collection of evidence as the physical aspect. In earlier class lectures, we learned forensic scientists consisted of chemists, who analyze and detect the presence of controlled substances in the body, and biologists, who use serology to detect substances found in stains. Now the focus has turned to the identification of toxins and poisonous substances that...
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...Serology CRP * C-Reactive Protein * An acute phase reactant * A substance in the blood that increases as a response to an acute conditions such as infection, injury, tissue destruction * First discovered in patients that react with the C polysaccharide of pneumococcus * Produced by the Liver * Elevates during inflammation Procedure Significance * To check for inflammation * To determine if an anti-inflammatory drug is working * Normal levels does not always indicate no inflammation * SLE, Rheumatoid Arthritis, Scleroderma, Polymyositis and dermatomyositis can elicit no a negative result for CRP. Positive results can appear in: * Cancer * Connective tissue disease * Heart attack * Infection * Inflammatory bowel disease (IBD) * Pneumococcal pneumonia * Tuberculosis ASO * Anti-Streptolysin O * Antibody specific for Streptolysin O, a toxin produced by groups A, C and G streptococci * Streptolysin O causes beta hemolysis * ASO titer is used to determine a recent streptococcal infection * Measured in Todd units * 6 hour fasting is needed before blood is to be extracted Procedure Significance * To confirm streptococcal infection * Used as diagnosis or confirmatory test for rheumatism or scarlet fever * ASO titer is based on the neutralization of the streptococcal enzyme streptolysin O. * Some antibiotics and corticosteroids may decrease ASO antibody levels...
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...Immigration Health Programmes (address below) for Citizenship and Immigration Canada. Generally, applicants for immigration are only given Section A of the Medical Report form and do not receive Sections B to E. You should have copies of the form IMM 5419, which contains Sections B to E. If you do not, please write to the Director of Immigration Health Programmes, at the address below, for information on how to obtain the form IMM 5419 on CD ROM. 2. Procedures for medical examination: (a) The person presenting this form requires the following medical tests: - complete medical examination - for persons of all ages - chest x-ray and report - for persons 11 years of age and over - urinalysis - for persons 5 years of age and over - syphilis serology - for persons 15 years of age and over - HIV testing should be performed on all applicants who are 15 years of age and over, as well as on those children who have received blood or blood products, have a known HIV positive mother, or are potential adoptees (b) The results of the examination must be reported on form IMM1017 AND form IMM 5419. Form IMM 1017 + Form IMM 5419 = one Medical Report One complete Medical Report is required for each person examined. (c) Actual laboratory reports are required. 3. Procedures for x-ray: (a) The radiographer will assure himself as to the identity of the person to be...
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