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DIAGNOSTIC PROCEDURES – STREPTOCOCCUS PYOGENES NEISSERIA GONORRHOEAE INFLUENZA VIRUS HUMAN IMMUNODEFICIENCY VIRUS.

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Presentation on theme: "DIAGNOSTIC PROCEDURES – STREPTOCOCCUS PYOGENES NEISSERIA GONORRHOEAE INFLUENZA VIRUS HUMAN IMMUNODEFICIENCY VIRUS."— Presentation transcript:

1 DIAGNOSTIC PROCEDURES – STREPTOCOCCUS PYOGENES NEISSERIA GONORRHOEAE INFLUENZA VIRUS HUMAN IMMUNODEFICIENCY VIRUS

2 Streptococcus pyogenes – GROUP A – BETA HEMOLYTIC STREP – GAS – GABHS – GRAM POSITIVE COCCI IN CHAINS PHARYNGITIS – SORE THROAT – TONSILITIS COLONIZATION OF PHARYNX, TONSILS, LARYNX (LESS OFTEN) ALSO: IMPETIGO, ENDOCARDITIS, MENINGITIS TRANSMISSION – CLOSE CONTACT, 1-3 DAYS KIDS – 12% ARE CARRIERS WITH NO SYMPTOMS SYMPTOMS – SORE THROAT, FEVER >100.4 F, PUS ON TONSILS, SWOLLEN LYMPH NODES IN NECK (CERVICAL), NO COUGH 3-5 DAYS UNTREATED COMPLICATIONSACUTE RHEUMATIC FEVER, GLOMERULONEPHRITIS, PERITONSILAR ABSCESS, CERVICAL LYMPHADENITIS, MASTOIDITIS (AIR CELLS OF SKULL BEHIND EAR),

3 DIAGNOSIS: CLINICAL SYMPTOMS – RAPID ANTIGEN DETECTION TEST – RADT, 15 MIN 70% SENSITIVITY, 95% SPECIFICITY POSITIVE RADT – TREAT, NO CULTURE NEEDED NEGATIVE RADT – THROAT SWAB CULTURE – THROAT SWAB, 90-95% SENSITIVITY, 1-2 DAYS GRAM POSITIVE STREPOCOCCI ON MEDIUM CONTAINING BLOOD – HEMOLYSIS, CLEAR STREPTOCOCCAL RADT GABHS – CELL WALL POLYSACCHARIDE IS ANTIGEN A THROAT SWAB TO COLLECT CELLS EXTRACT POLYSACCHARIDE MIX WITH ENZYME-LINKED ANTI-GROUP A STREP ANTIBODIES ANTIBODIES BIND IF GAS IS PRESENT ADD CHROMOGENIC SUBSTRATE OF ENZYME COLOR – GAS (GROUP A STREP) WAS PRESENT IN SWAB

4 TREATMENT - PENICILLIN V – REDUCES: DURATION ~ 1 DAY (2-4 DAY DURATION) CONTAGION RISK OF COMPLICATIONS – ERYTHROMYCIN, CEPHALOSPORINS INCIDENCE – PHARYNGITIS 11 MILLION/YR IN US – MOSTLY VIRAL % - STREP THROAT ECONOMIC COSTS: ~$350 MILLION FOR CHILDREN/YR IN US

5 Neisseria gonorrhoeae – GRAM NEGATIVE DIPLOCOCCUS - GONOCOCCUS GONORRHEA – COLONIZATION OF MUCOUS MEMBRANES – URETHRA/MALES, CERVIX/FEMALES, EYE (CONJUNCTIVITIS), RECTUM, THROAT TRANSMISSION – CONTACT 2-8 DAYS – MALES; 1-3 WEEKS – FEMALES SYMPTOMSMALES – PAINFUL URINATION; WHITE-YELLOW DISCHARGE 10-15% NO SYMPTOMS FEMALES – MILDER; SOME VAGINAL DISCHARGE 35-80% NO SYMPTOMS NEWBORNS – CONJUNCTIVITIS/BLINDNESS COMPLICATIONS MALES – EPIDIDYMIS, PROSTATE INFECTIONS; URETHRA SCARRING FEMALES- INFECTION OF UTERUS, FALLOPIAN TUBES, PID, ECTOPIC PREGNANCY, INFERTILITY BACTEREMIA, ARTHRITIS, ENDOCARDITIS, MENINGITIS

6 DIAGNOSIS: GRAM STAIN – SWAB OF DISCHARGE, URETHRA (MALES) CERVIX(FEMALES) GRAM NEGATIVE DIPLOCOCCI WITHIN NEUTROPHILS CULTURE DNA AMPLIFICATION TEST – PRODUCTION OF FLUORESCENT DNA SPECIFIC FOR N. GONORRHOEAE AND SEPARATELY ALSOFOR CHLAMYDIA TRACHOMATIS TREATMENT:CEFTRIAXONE (CEPHALOSPORIN – BETA LACTAMS WHICH INHIBIT PEPTIDOGLYCAN SYNTHESIS/ PLUS AZITHROMCIN (MACROLIDE) – PROTEIN SYNTHESIS OR DOXYCLINE (TETRACYCLINE) PROTEIN SYNTHESIS ALSO CONTROLS CHLAMYDIA INCIDENCE:~700,000 NEW CASES/YR IN US. ~106 MILLION/YR IN BIG WORLD

7 INFLUENZA VIRUS – NEGATIVE STRAND, SS-RNA, 8 SEGMENTS, ~10 KB VIRAL RNA-DEPENDENT RNA POLYMERASE IN VIRIONS HEMAGLUTININ AND NEURAMINIDASE ON SURFACE INFECTS EPITHELIUM (HEMAGLUTININ STICKS TO SIALIC ACID) NOSE, THROAT, LUNGS, BIRD INTESTINE, SIALIC ACID = N-ACETYL NEURAMINIC ACID = MONOSACCHARIDE ON ALL CELL SURFACES ENDOCYTOSIS; CORE PROTEINS, RNA AND RNA-DEPENDENT RNA POL ENTER NUCLEUS, TRANSCRIPTION, TRANSLATION, ASSEMBLY OF VIRIONS IN NUCLEUS; VIRAL PROTEINS HYDROLYZE HOST mRNA, INHIBIT HOST TRANSLATION BUDDING THRU CYTOPLASMIC MEMBRANE, ACQUIRING HEMAGLUTININ, NEURAMINIDASE (ENZYME WHICH CLEAVES HOST SIALIC ACID ON CELL SURFACT TO RELEASE VIRUS) HOST CELLS DIE

8 ANTIGENIC DRIFT AND SHIFT DRIFT RNA-DEPENDENT RNA POL – ONE MUTATION/10 KB REPLICATED, THEREFORE – MOST NEW VIRUS PARTICLES ARE MUTANTS, HAVE ALTERED SURFACE PROTEINS, SLOW CHANGE IN SURFACE ANTIGENS OVER TIME SHIFT WHEN TWO DIFFERENT FLU VIRUS PARTICLES INFECT SINGLE CELL, RNA SEGMENTS MIX (RE-ASSORT) RAPID ANTIGEN CHANGES ACQUIRE NEW HOST SPECIES; ESCAPE PROTECTIVE IMMUNITY PANDEMICS EMERGE

9 TRANSMISSION – FROM ~1 DAY BEFORE SYMPTOMS – ~7 – 14 DAYS CONTACT – DIRECT – HANDSHAKE > THEN TO EYES, MOUTH - DROPLETS - SNEEZING MUCUS INTO EYES, NOSE, MOUTH ~40,000 DROPLETS/SNEEZE - INDIRECT-CONTAMINATED SURFACE > THEN TO EYES, MOUTH VEHICLES – DROPLET NUCLEI – 0.5 – 5 MICROMETER DIAMETER TRAVEL LONG DISTANCES SYMPTOMS – MIX OF COMMON COLD AND PNEUMONIA (INFLAMMATION LUNG AFFECT ALVEOLI -AIR SACS-, DIFFICULTY BREATHING) CHILLS, FEVER, CONGESTION, FATIGUE, BODY ACHES/PAIN 33% WITH FLU = ASYMPTOMATIC COMPLICATIONS – SECONDARY VIRAL/BACTERIAL PNEUMONIA DEATH

10 DIAGNOSIS - SYMPTOMS (DIFFICULT; COLD & PNEUMONIA) VIRUS REPLICATION IN TISSUE CULTURE (3-10 DAYS) NASOPHARYNGEAL – THROAT SWAB, NASOPHARYNGEAL, BRONCHIAL WASH NASAL/ENDOTRACHEAL ASPIRATE (MATERIAL DRAWN OUT) SPUTUM REVERSE TRANSCRIPTION PCR (HOURS) DETECTS INFLUENZA RNA FLUORESCENT ANTIBODIES TO INFLUENZA ANTIGENS (HOURS) RAPID INFLUENZA DIAGNOSTIC TESTS (RIDT) MIN DETECTS INFLUENZA ANTIGENS A OR B ENZYME-LINKED ANTI-INFLUENZA ANTIBODIES TREATMENT - NEURAMINIDASE INHIBITOR TAMIFLU EFFECTIVENESS QUESTIONABLE – REDUCES DURATION BY 13% CHILDREN - AVOID ASPIRIN - CAN CAUSE REYES’S SYNDROME POTENTIALLY FATAL BRAIN/LIVER DISEASE MOSTLY OF KIDS TAKING ASPIRIN AFTER VIRAL INFECTION

11 VACCINES - TRIVALENT – PURIFIED FLU ANTIGENS TWO TYPE B, ONE TYPE A NEW VACCINES PREPARED YEARLY – NEW STRAINS REPLACE OLD ONES ANTIGENIC SHIFTS – NEW STRAINS WITH SLIGHTLY DIFFERENT ANTIGENS WORLD HEALTH ORGANIZATION – TRYS TO PREDICT STRAINS WHICH WILL CAUSE SEASONAL EPIDEMICS SEASONAL EPIDEMICS – 3-5 MILLION SEVERE CASES/YR; 500,00 DEATHS/YR WORLD 200,000 HOSPITALIZATONS/YR; ~3000 – 49,000 DEATHS/YR USA PANDEMICS SEVERE SYMPTOMS – ,000 MILLION DEATHS/WORLD SPANSH FLU, 2-20% MORTALITY, 50% INFECTION RATE 2.5-5% OF WORLD POPULATON H1N1 [HEMAGLUTININ 1, NEURAMINIDASE 1]

12 AVIAN STRAIN SPREAD TO HUMANS – 1990s H5N1 – ANTIGENIC SHIFT DOES NOT SPREAD EASILY BETWEEN HUMANS A/H1N STRAIN EVOLVED FROM HUMAN, PIG AND BIRD GENES LOWER MORTALITY RATE THAN SEASONAL FLU 18,000 DEATHS

13 HUMAN IMMUNODEFICIENCY VIRUS – ENVELOPED, SS RNA ~9000 BASE RETROVIRUS, FUSION, RELEASE OF NUCLEOCAPSID CYTOPLASM, REVERSE TRANSCRIPTION AND RNA HYDROLYSIS PRODUCES DS DNA ENTRY NUCLEUS, INTEGRATION AS PROVIRUS, ACTIVATION, TRANSCRIPTION PRODUCING VIRAL RNA (SERVES AS MESSENGER & PROGENY CHROMOSOMES, VIRAL PROTEINS PRODUCED, VIRIONS ASSEMBLED, HIV PROTEINS INSERTED INTO CYTOPLASMIC MEMBRANE BUDDING RELEASES PROGENY VIRUSES, DEATH OF HOST CELL ACQUIRED IMMUNODEFICIENCY SYNDROME – AIDS TRANSMISSION -SEXUAL INTERCOURSE = MAJOR MODE HIV FREE VIRUS AND INSIDE INFECTED LYMPHOCYTES BLOOD, SEMEN, VAGINAL SECRETIONS, BREAST MILK, PRE-EJACULATE FLUID INFECTS CD4+ T LYMPHOCYTES, MACROPHAGES, DENDRITIC CELLS CORECEPTORS – CCR5 AND CXCR4 (CHEMOKINE RECEPTORS)

14 GENETIC VARIABILITY - 9,000 BASE GENOME, 3 X 10e-5 MUTATIONS/NUCLEOTIDE POLYMERIZED, ~1 MUTATION FOR EVERY 3 MOLECULES RNA 1 X 10e10 NEW PROGENY VIRUSES/DAY/PERSON ~3 X 10e9 MUTATIONS IN HIV POPULATION/DAY/PERSON AIDS: GRADUAL LOSS OF CD4+ T LYMPHOCYTES VIRUS KILLING; ATTACK BY IMMUNE CYTOTOXIC T LYMPHOCYTES OPPORTUNISTIC INFECTIONS – LIFE THREATENING CANCER (T CELL LYMPHOMA) PATHOGENS

15 DIAGNOSIS – ELISA – ANTI-HIV ANTIBODIES WINDOW – TIME FROM INFECTION TO TIME DETECTABLE ANTIBODIES MOSTLY ~ 30 DAYS; RANGE 3 WEEKS – 3 MONTHS INFECTIOUS !!! DILUTED SERUM APPLIED TO STRIP LOADED WITH HIV ANTIGENS APPLY ENZYME-LINKED ANTIBODIES TO HUMAN IgG ADD CHROMOGENIC SUBSTRATE FOR ENZYME LOOK FOR COLOR – INDICATES HIV ANTIBODIES PRESENT HIGH SENSITIVITY – LOW SPECIFICITY POSITIVE RESULT IS FOLLOWED BY WESTERN BLOT TO CONFIRM WESTERN- DETECTS ANTI-HIV ANTIBODIES HIV PROTEINS – 3 MAJOR – SEPARATED BY MW ON ELECTROPHORESIS GEL > MEMBRANE FLOOD MEMBRANE WITH DILUTED PATIENT SERUM (HIV +/-) PROBE WITH ENZYME-LINKED ANTI-HUMAN IgG ANTIBODIES ADD SUBSTRATE > RECORD ENZYME REACTION > INDICATES HIV+

16 ORAQUICK RAPID HIV TEST – HOME USE – TESTS ANTI-HIV ANTIBODIES IN MUCOUS SECRETION – UNDER THE LIP FALSE NEGATIVES ~5 PERCENT NAT -NUCLEIC ACID BASED TEST RNA EXTRACTED FROM PLASMA REVERSE TRANSCRIPTASE PCR CONVERTS RNA TO DNA DETECT PRODUCT DETECTION LIMIT = 50 MOLECULES RNA/ML PCR MIX, BUT PLASMA DILUTED 1:20 DETECTION LIMIT IN PLASMA = 1000 MOLECULES/ML WINDOW - ~17 DAYS NEWBORNS – MOTHER HIV+ WILL HAVE ANTI-HIV ANTIBODIES MUST CHECK BY PCR VIRAL RNA


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