Intracellular, Virology, Viral Specimens, Mycoplasma/Ureaplasma
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| What is unique to the Chlamydiaceae reproductive cycle? | Elementary bodies and reticulate bodies are unique to the Chlamydiacea reproductive cycle
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| how are specimens for Chlamydiacea transported or stored? | Must be kept cold and transported in special-antibiotic containing media. Refrigerate on receipt and freeze at -70 C if unable to process within 24 hours
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| How do labs ID Chlamydiaceae | Labs mostly ID Chlamydiaceae using Immunofluorescence and Nucleic Acid Assays
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| What is seen on an Iodine stain of Chlamydiacea trachomatis? | Compact inclusions in the epithelial cells. Stain darkly and contain glycogen
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| What is seen on an Iodine stain of Chlamydiacea psittaci and Chlamydiacea pneumoniae? | diffuse inclusions that do not stain.
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| How do all elementary bodies in Chlamydiacea organisms appear on Giemsa stain? | reddish-purple
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| How do all reticulate bodies in Chlamydiacea organisms appear on Giemsa stain? | Blue
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| What nucleic acid assays are used for Chlamydiacea trachomatis? | DNA probes and PCR are used for Chlamydiaciae trachomatis and PCR is the most sensitive being able to detect as few as 10 EB in a specimen.
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| What serology is used for Chlamydiacea? | complement fixation, immunofluorescence, ELISA
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| Sx of Chlamydia (non-gonococcal urethritis)/ | Anogenital discharge, burning with urination and/or rectal pain. Men do not have symptoms as much as women.
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| Complications of Chlamydia? | Males do not often have complications but women which causes permanent damage to the fallopian tubes, uterus, and surrounding tissues and leading to ectopic pregnancy or infertility
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| Tx of Chlamydia | Tx w/ the antibiotics azithromycin or doxycycline typically
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| Trachoma is the most common cause of what? | preventable blindness of infectious origin.
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| What ocular condition can be caused by a Neonatal infection with Chlamydiacea? | Inclusion conjunctivitis
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| What are the symptoms of inclusion conjunctivitis? | redness of the eyes, swelling of the eyelids, and discharge of pus and likely to appear 5-12 days after birth.
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| What are the symptoms of Lymphogranuloma venereum? | initially a small, painless, vesicular lesion results in a draining ulcer that heals spontaneously. 2-6 weeks later: chills, fever, and lymphadenopathy.
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| How is Lymphogranuloma venereum diagnosed? | genital and lymph node specimens are tested for Chlamydiaceae trachomatis by culture, immnofluoresence or nucleic acid amplification.
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| How do humans acquire the zoonotic agent Psittacosis? | inhalation of infection, organism is deposited in alveoli and may be ingested by macrophages and transported to lymph nodes where they may be disseminated via RES through the body
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| Sx of psittacosis | Fever, chills, fatigue, headache, non-productive cough.
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| Tx of psittacosis? | tetracycline is the drug of
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| Atypical pneumonia transmission | person-to-person via respiratory droplets.
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| What age group is at greatest risk for atypical pneumonia? | adolescents, young adults, and older adults at greatest risk.
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| Sx of atypical pneumonia? | usually mild, most common complaint is a persistent cough
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| Tx of atypical pneumonia? | tetracycline, doxycycline, macrolides, fluoroquinolones, or erythromycin
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| Causative agent of Q fever? | Coxiella burnetii
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| What kind of lab must handle any Coxiella burnetii suspected specimens? | BSL-III
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| Sx of Q fever? | acute sx of fever, severe headache, malaise, myalgia, NVD, non-productive cough. chronic sx of pneumonia, hepatitis, myocarditis, and CNS complications
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| Mode of transmission of Q fever? | primarily spread via inhalation with a very low infectious dose.
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| Which variant is the infectious form of Coxiella burnetii? | Small cell variants are infectious
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| Dx of Q fever based on | low PLT, normal WBC, increased liver enzymes. PCR early in illness, IFA is the gold standard for confirmation. A 4-fold rise in titer seen in IFA between acute and convalescent is diagnostic
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| TX of Q fever? | Doxycycline is the treatment
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| Prognosis of Q fever? | Rarely fatal, <2% of patients
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| What safety level is required for virology? | Most labs just need BSL 2, SARS, Influenza cell cultures, Hemorrhagic fevers, and smallpox would require BSL 3 or 4.
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| Time of specimen collection for virology? | Ideally, no more than 3-5 days after onset of symptoms, but as soon as possible
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| What are the requirements for virol specimen collection materials? | Sterile, secure, leak-proof, non-breakable, able to withstand freezing/thawing. Swabs must use dacron, rayon, or other polyester fiber swabs with plastic or aluminum shaft.
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| What is in transort media? | antibiotics and antifungals to prevent overgrowth of other microorganisms and components to preserve infectivity.
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| What temperature should viral specimens be stored at for transport? | 2-8 celsius on wet ice or cold packs. if longer than 48 hours delay, freeze at -70C, otherwise do not freeze.
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| What is a cell culture? | metabolically active mammalian cells grown in vitro that are treated w/ a chemical agent to dissociate them and suspended in a growth medium and dispensed into tubes or flasks. The cells settle to one side and form a monolayer.
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| Shell vial culture setup? | diploid cells are often used. monolayer grows on coverslip in small glass/plastic shell vial. specimen inoculated into shel vial, centrifuged for 1 hr and incubated at 35 C for 24-48 hours and viral antigens detected by FITC-labeled monoclonal antibodies
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| Monolayer cell culture setup? | innoculate w/ 0.2-0.2 mL processed specimen, incubate 12-24 hrs @34 C in aerobic environment. Hold in roller drum. Refresh maintenance medium 1-2x/week during incubation. monitor > 3x/week for visible color change of medium or viral prolif microscopically
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| Why do scientists say viruses are not alive? | Do not contain BOTH DNA and RNA, do not divide, do not produce energy, lack building blocks for protein and nucleic acid production, require a host cell to multiply.
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| What are the parts of a virion? | nucleic acid (DNA or RNA) with a protein coat and tend to be either enveloped or nonenveloped.
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| What are the steps of viral replication? | 1. attachment 2. penetration 3. uncoating 4. macromolecular synthesis/transcription 5. macromolecular synthesis transcription, protein synthesis, and replication 6. Lysis and release or budding and release.
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| Attachment step of viral replication | virus attaches to surface protein or polysaccharide of host celll
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| Penetration step of viral replication | entier virus enters cell through fusion/endocytosis
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| uncoating step of viral replication | viral nucleic acid released from capsid and virus now controls the cell.
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| biosynthesis or eclipse step of viral replication | virus induces host cell machinery to synthesize components for viral replication, may occur after dormancy.
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| assembly/maturation step of viral replication | newly synthesized viral components are assembled into a new virion
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| release step of viral replication | escape the host cel by lysis which destroys the cell or budding to form enveloped virus. Either way this can then infect new host cells
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| Which viruses are DNA viruses? | Herpes simplex viruses and family are all DNA, Smallpox, HPV, Adenovirus, and Hep B are all DNA as well.
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| What virus causes cold sores/fever blisters? | HSV1 (Herpes Labialis)
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| What virus causes genital herpes? | HSV-2 (Herpes Genitalis)
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| What causes chicken pox? | Varicella
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| What causes shingles? | Herpes Zoster
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| Why does chicken pox sometimes lead to shingles? | Varicella-Zoster Virus (HHV-3) is the same virus, herpes zoster is the reactivation of VZV in the nerves.
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| What causes infectious mononucleosis (mono) | Epstein-Barr Virus (HHV-4)
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| What causes Roseola (or 6th disease) which has a 3-4 day fever followed by maculopapular rash for 1-2 days? | HHV-6 or HHV-7
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| What fall into the herpesviridae family? | Herpes simplex viruses (HSV-1,-2), Varicella Zoster Virus (HHV-3), Epstein-Barr Virus (HHV-4), Cytomegalovirus (HHV-5), Human Herpes Viruses (HHV-6, -7, -8)
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| How many strains of Human Papillomavirus (HPV) are there? | 70 different viruses
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| What viruses fall under the Picornaviradae family? | Rhinovirus, enterovirus (Polio, coxsackie A and B, Echoviruses, Enteroviruses),
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| What virus causes the common cold? | Rhinovirus
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| What causes Hand, foot, and mouth disease? | Coxsackie A16 virus
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| What causes Hemorrhagic conjunctivitis? | Coxsackie A24 virus
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| What causes encephalitis? | Arboviruses which are ARthropod-BOrn VIRUSes
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| SARS belongs to what family? | Coronaviridae
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| What causes the acute upper respiratory illness of children that has "croup" as a symptom? | parainfluenza
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| What causes cold-like symptoms in healthy adults but is more serious in elderly and infants? | Respiratory Syncytial Virus (RSV)
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| What causes Measles? | Rubeola
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| What viruses fall under the paramyxoviridae family? | Parainfluenza, Respiratory Syncytial Virus (RSV), Rubeola, Mumps
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| What does Mumps virus cause? | swollen tender salivary (parotid) glands and fever
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| What causes German measles? | Rubella
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| What viruses cause viral gastroenteritis? | Norovirus and Rotavirus
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| Transmission of norovirus and rotavirus? | Norovirus is transmited via fecal-oral, airborne or fomite transmission (esp. hospitals, cruise ships, nursing homes, etc.) Rotavirus is most commonly a concern for infants
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| How are mycoplasma and ureaplasma different from other bacteria? | they do not possess a cell wall but a membrane. This makes gram staining not work.
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| Are Mycoplasma pneumoniae, M. hominis, ureaplasma urealyticum, and M. genitalium normal flora? | No
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| How are Mycoplasma pneumoniae, M. hominis, ureaplasma urealyticum, and M. genitalium transmitted? | direct sexual contact, transplant tissue from infected donors, mother to fetus at birth, or (M. pneumoniae only) respiratory secretions.
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| how does Mycoplasma pneumoniae manifest in school age children? | Upper resp Infection with mild, non-specific symptoms
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| how does Mycoplasma pneumoniae manifest in adolescents and young adults? | Lower resp infection. Flu-like symptoms and can include rash, arthritis, encephalitis, myocarditis, pericarditis, and hemolytic anemia as complications.
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| How does Mycoplasma hominis manifest? | systemic infections in neonates resulting from mother-to-fetus transmission, invassive disease in immunosuppressed, GU infections, also associated with bacterial vaginosis
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| How does Ureaplasma urealyticum manifest? | same as M. hominis as well as an association w/ chronic lung disease
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| How does Mycoplasma genitalium manifest? | nongonococcal urethritis in men and possible cervicitis and endometritis in females. mother-to-fetus transmission possible but unknown significance.
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| What stains are used for the Mycoplamsa and Ureaplasma species? | Acridine orange is useful but nonspecific and used because gram stain would not be useful due to the lack of a cell wall.
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| What media is used for Mycoplasma and Ureaplasma species growth? | The organism needs sterols for development of membranes, yeast extract gives them growth factors, pH indicator to detect growth, penicillin to minimize overgrowth of other bacterias, and metabolic substrate for the suspected species.
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| What does Mycoplasma pneumoniae metabolize? | Glucose
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| What does Mycoplasma hominis metabolize? | Arginine
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| What does Ureaplasma urealyticum metabolize? | Urease
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| What does Mycoplasma genitalium metabolize? | Glucose
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| how are the colonies of Mycoplasma and Ureaplasma observed on the agar? | Using stero microscopy
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| What is the colonial appearance of Mycoplasma pneumoniae? | 50-500 um diameter, spherical, granular yellowish colonies
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| What is the colonial appearance of Mycoplasma hominis? | 20-300 um fried egg colonies
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| What is the colonial appearance of Ureaplasma urealyticum? | 15-60 um bird's nest, small, granular colonies
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| What is the colonial appearance of Mycoplasma genitalium? | <1 um diameter fried egg colonies
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| What is the developmental cycle of Chlamydiaciae? | Elementary bodies enter an epithelial cell The elementary bodies then differentiate into reticulate bodies (metabolically active and noninfective chlamydiaciae cells) These reticulate bodies then divide, revert to elementary bodies and evacuate the cell.
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| How are mycoplasma and ureaplasma transported? | Bodie fluids and tissue must either be submitted within 1 hour of collection or held for up to 24 hours @ 4C. Swabs must be placed immediately into transport media up to 24 hrs @ 4C. >24 hr, freeze @ -70C
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| What special requirement is there for tissues when mycoplasma or Ureaplasma are suspected? | The tissue is diced with a scalpel instead of grinding with mortar and pestle.
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