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Micro Exam 4

Intracellular, Virology, Viral Specimens, Mycoplasma/Ureaplasma

What is unique to the Chlamydiaceae reproductive cycle? Elementary bodies and reticulate bodies are unique to the Chlamydiacea reproductive cycle
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
How do labs ID Chlamydiaceae Labs mostly ID Chlamydiaceae using Immunofluorescence and Nucleic Acid Assays
What is seen on an Iodine stain of Chlamydiacea trachomatis? Compact inclusions in the epithelial cells. Stain darkly and contain glycogen
What is seen on an Iodine stain of Chlamydiacea psittaci and Chlamydiacea pneumoniae? diffuse inclusions that do not stain.
How do all elementary bodies in Chlamydiacea organisms appear on Giemsa stain? reddish-purple
How do all reticulate bodies in Chlamydiacea organisms appear on Giemsa stain? Blue
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.
What serology is used for Chlamydiacea? complement fixation, immunofluorescence, ELISA
Sx of Chlamydia (non-gonococcal urethritis)/ Anogenital discharge, burning with urination and/or rectal pain. Men do not have symptoms as much as women.
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
Tx of Chlamydia Tx w/ the antibiotics azithromycin or doxycycline typically
Trachoma is the most common cause of what? preventable blindness of infectious origin.
What ocular condition can be caused by a Neonatal infection with Chlamydiacea? Inclusion conjunctivitis
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.
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.
How is Lymphogranuloma venereum diagnosed? genital and lymph node specimens are tested for Chlamydiaceae trachomatis by culture, immnofluoresence or nucleic acid amplification.
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
Sx of psittacosis Fever, chills, fatigue, headache, non-productive cough.
Tx of psittacosis? tetracycline is the drug of
Atypical pneumonia transmission person-to-person via respiratory droplets.
What age group is at greatest risk for atypical pneumonia? adolescents, young adults, and older adults at greatest risk.
Sx of atypical pneumonia? usually mild, most common complaint is a persistent cough
Tx of atypical pneumonia? tetracycline, doxycycline, macrolides, fluoroquinolones, or erythromycin
Causative agent of Q fever? Coxiella burnetii
What kind of lab must handle any Coxiella burnetii suspected specimens? BSL-III
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
Mode of transmission of Q fever? primarily spread via inhalation with a very low infectious dose.
Which variant is the infectious form of Coxiella burnetii? Small cell variants are infectious
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
TX of Q fever? Doxycycline is the treatment
Prognosis of Q fever? Rarely fatal, <2% of patients
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.
Time of specimen collection for virology? Ideally, no more than 3-5 days after onset of symptoms, but as soon as possible
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.
What is in transort media? antibiotics and antifungals to prevent overgrowth of other microorganisms and components to preserve infectivity.
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.
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.
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
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
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.
What are the parts of a virion? nucleic acid (DNA or RNA) with a protein coat and tend to be either enveloped or nonenveloped.
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.
Attachment step of viral replication virus attaches to surface protein or polysaccharide of host celll
Penetration step of viral replication entier virus enters cell through fusion/endocytosis
uncoating step of viral replication viral nucleic acid released from capsid and virus now controls the cell.
biosynthesis or eclipse step of viral replication virus induces host cell machinery to synthesize components for viral replication, may occur after dormancy.
assembly/maturation step of viral replication newly synthesized viral components are assembled into a new virion
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
Which viruses are DNA viruses? Herpes simplex viruses and family are all DNA, Smallpox, HPV, Adenovirus, and Hep B are all DNA as well.
What virus causes cold sores/fever blisters? HSV1 (Herpes Labialis)
What virus causes genital herpes? HSV-2 (Herpes Genitalis)
What causes chicken pox? Varicella
What causes shingles? Herpes Zoster
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.
What causes infectious mononucleosis (mono) Epstein-Barr Virus (HHV-4)
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
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)
How many strains of Human Papillomavirus (HPV) are there? 70 different viruses
What viruses fall under the Picornaviradae family? Rhinovirus, enterovirus (Polio, coxsackie A and B, Echoviruses, Enteroviruses),
What virus causes the common cold? Rhinovirus
What causes Hand, foot, and mouth disease? Coxsackie A16 virus
What causes Hemorrhagic conjunctivitis? Coxsackie A24 virus
What causes encephalitis? Arboviruses which are ARthropod-BOrn VIRUSes
SARS belongs to what family? Coronaviridae
What causes the acute upper respiratory illness of children that has "croup" as a symptom? parainfluenza
What causes cold-like symptoms in healthy adults but is more serious in elderly and infants? Respiratory Syncytial Virus (RSV)
What causes Measles? Rubeola
What viruses fall under the paramyxoviridae family? Parainfluenza, Respiratory Syncytial Virus (RSV), Rubeola, Mumps
What does Mumps virus cause? swollen tender salivary (parotid) glands and fever
What causes German measles? Rubella
What viruses cause viral gastroenteritis? Norovirus and Rotavirus
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
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.
Are Mycoplasma pneumoniae, M. hominis, ureaplasma urealyticum, and M. genitalium normal flora? No
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.
how does Mycoplasma pneumoniae manifest in school age children? Upper resp Infection with mild, non-specific symptoms
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.
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
How does Ureaplasma urealyticum manifest? same as M. hominis as well as an association w/ chronic lung disease
How does Mycoplasma genitalium manifest? nongonococcal urethritis in men and possible cervicitis and endometritis in females. mother-to-fetus transmission possible but unknown significance.
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.
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.
What does Mycoplasma pneumoniae metabolize? Glucose
What does Mycoplasma hominis metabolize? Arginine
What does Ureaplasma urealyticum metabolize? Urease
What does Mycoplasma genitalium metabolize? Glucose
how are the colonies of Mycoplasma and Ureaplasma observed on the agar? Using stero microscopy
What is the colonial appearance of Mycoplasma pneumoniae? 50-500 um diameter, spherical, granular yellowish colonies
What is the colonial appearance of Mycoplasma hominis? 20-300 um fried egg colonies
What is the colonial appearance of Ureaplasma urealyticum? 15-60 um bird's nest, small, granular colonies
What is the colonial appearance of Mycoplasma genitalium? <1 um diameter fried egg colonies
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.
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
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.
Created by: wulfmannwarrior