| Organism/ Infection | Specimen/ Site | Kit(s) Available at MHD - Laboratory (Specimen Handling) | Procedure | General Information |
|---|
BACTERIOLOGY |
Bacterial enteric pathogen infections: Salmonella, Shigella, Yersinia, Campylobacter
E. coli 0157:H7 Form Number:
H-429A 989 R3/97 |
Stool specimen |
Enteric Pathogen Kit. Includes: vial of Carey-Blair (red) transport medium, plastic stool container, lab requisition, and instruction sheet. Transportation: Transport specimen in a plastic zip-lock bag on wet ice, with the lab requisition in the outer pouch.
Storage: Refrigerate the specimen and unused Carey-Blair transport medium.
Turnaround Time: Negative: 2 days Presumptive positive: 2-3 days Confirmed: 3-7 days |
Appropriate media are inoculated directly with the specimen and also after an enrichment step. Suspected organisms are identified using biochemical tests and serological typing. |
From 90 stool specimens processed in 2003, 12.0% were positive for enteropathogenic bacteria. Salmonella was the most commonly isolated enteric pathogen. |
| Bordetella pertussis(whooping cough) Form Number:
H-429A 989 R3/97 |
Nasopharyngeal |
Pertussis Kit. Includes: Plate and tube of Regan-Lowe medium, 2 ringed slides, lab requisition and instruction sheet. Keep refrigerated until use (DO NOT FREEZE)
Transportation: Deliver as soon as possible. If delay is expected, incubate inoculated culture plate and transport medium tube at 35oC until delivery is possible.
Turnaround Time: DFA same day. Culture 3-7 days. |
DFA: Smear is stained with fluorescent antibody and examined using fluorescent microscopy
Culture: Regan-Lowe plates are inoculated directly with specimen or from transport medium and incubated at 35oC and examined daily. Characteristic colonies confirmed by direct fluorescent antibody (DFA). |
Twenty eight cases of
B. pertussis were detected by the Milwaukee HealthDepartment Laboratories.
Majority of the cases were detected during the months of August through December which was consistent with the previous year's observations (1987-2002).
 Detection of Pertussis by Culture or DFA Test (1983-2003)
Culture Confirmed Pertussis Cases by Month,
Cumulative (1987-2003) |
| Referred culture Identification Form Number
H-429A 989 R3/97 |
Pure culture of isolate submitted in tube or plate |
No kit is required.
Transportation: Pure culture in a tube or plate. Transport at room temperature.
Extended storage at 4oC. Turnaround Time: 7-21 days |
Various biochemical tests and cellular fatty acid analysis are used for identification. |
624 isolates were identified in 2003. |
| Strep Throat Form Number
H-429A 12/90 |
Throat |
Culturette swab.
Transportation: 25o-30oC. Storage: 25o-30oC.
Turnaround Time: 24 hours. |
Cultured on blood agar plate with 0.04 U bacitracin disc. |
25.0% of specimens received were positive for Group A Beta-strep in 2003. |
| Tuberculosis Form Number
H-432 R12/99 |
Sputum |
Sputum collection kit. Includes: sterile 50 ml. collection tube and a leak-proof container). Transportation: Transport on wet ice.
Storage: Refrigerator temperature.
Turnaround Time: 3-6 Weeks. |
Concentrated specimens are examined microscopically after staining and plated onto solid media. Portions are inoculated into Bactec vials. Identification by biochemical testing and DNA probe. Susceptibility testing performed upon request. |
The most common isolates in 2003 were:
M. avium complex
Mycobacterium tuberculosis
|
| Legionella Evaluation (Culture/DFA) Form Number
H-429 989 R3/97 |
Respiratory specimens: Sputum, BAL, Bronchial wash,
Lung tissue, etc. |
Transportation: Deliver immediately to lab on wet ice or refrigerate at 4-8oC for up to 24 hours.
For extended storage, samples may be frozen at -20oC.
Turnaround Time: DFA: Same day.
Culture: 3 to 14 days. |
DFA - Monoclonal antibodies are used to identify all serogroups of Legionella pneumophila.
Portions of untreated and acid treated specimen are plated with selective and non-selective BCYE media plates. Characteristic colonies are identified. |
The number of positive cases increases during August through October (1983-2003).
Legionella Testing: Total and Positive Patients (1983-2003) |
| Legionellosis (Diagnostic) Form Number
H-429 989 R3/97 |
Urine (random) |
Transportation: Transport specimen at 4-8oC.
Turnaround Time: Same day for specimens received before noon. For those received after noon, results available the next day. |
Legionella pneumophila Serogroup I antigen detected in urine by enzyme immunoassay (EIA). |
Legionella urine antigen was detected in 3 of 295 patients tested in 2003. |
| Legionellosis (Diagnostic) Form Number
H-429 989 R3/97 |
Serum; Acute and Convalescent collected 2-4 weeks apart. |
Transportation: Transport at 2-4oC or on wet ice.
Storage: Serum may be refrigerated at 2-4oC for several days. For extended
storage, freezing at -20oC is recommended.
Turnaround Time: 7-14 days |
Quantitative total antibody to L. pneumophila serogroups 1-6 detected using indirect immunofluorescent assay (IFA).
Positive serological confirmation based on a four-fold increase in titer between acute and convalescent sera. |
No serological positive case was detected in 2003. |
Legionella Environmental Sample
Form Number
H-429 989 R3/97 |
500 ml of water in sterile, leak-proof container |
Contact Laboratory before collection.
Turnaround Time: 3 to 10 days. |
Approximately 300 ml of water is filtered. Trapped microorganisms on the filter are recovered and inoculated on a culture media. |
Aquatic environmental samples are tested in an outbreak situation. |
| Organism/ Infection | Specimen/ Site | Kit(s) Available at MHD - Laboratory (Specimen Handling) | Procedure | Statistics (2003) |
|---|
MYCOPLASMA |
Mycoplasma
(Genital or Oral) Form Number
H-429A 989 R3/97 |
Genital swab, urine, biopsy specimen Sputum, BAL, throat or nasopharyngeal swab |
Transport For optimal recovery, specimens should be refrigerated and delivered within 6 hr. after collection. If this is not possible, specimen should be placed in appropriate transport media and frozen at -20oC for up to 2 weeks or at -70oC for longer term storage.
Turnaround Time: Genital: 2-7 days, Oral: 2-6 weeks |
Specimens are cultured using agar-broth technique. Identification is performed on the basis of biochemical reactions and hemagglutination test. |
More than 1478 specimens were tested for oral and genital Mycoplasma in 2003.
Resp Genital
(312 + 1166) = 1478 total)
Genital Mycoplasma Specimens (1985-2003) |
|
SEROLOGY |
|
| Syphilis Serology (VDRL) Form Number
H-429 989 R3/97 |
Serum or CSF |
Whole blood tubes without anticoagulant, serum or CSF, free from visible contamination.
Transportation: Transport at room temperature.
Turnaround Time: 24 hours. |
A non-specific screening test that uses a cardiolipin-based antigen is used to detect Reagin (an anti-lipid substance) in reactive serum or CSF specimens. |
9,530 specimens were screened in 2003 with a positivity rate of 4.6%. |
| FTA-ABS Form Number
H-429 989 R3/97 |
Serum |
Transportation: Serum specimens are transported as above NOTE: This test is not performed on CSF.
Turnaround Time: 1 to 4 days. |
Qualitative IFA procedure specific for Treponema pallidum antibodies using T. pallidum antigen (Reiters strain). |
Reactive VDRL tests are confirmed by FTA-ABS test.
312 specimens tested in 2003 with a positivity rate of 49.4%.
Total FTA-ABS and Reactive Specimens (1985-2003) |
|
MYCOLOGY |
|
| Ringworm/ dermatophytes Form Number
H-429 989 R3/97 |
Hair, Skin, Nails |
Collect scrapings from the infected site in a small clean envelope.
Transportation: Room temperature
Turnaround Time: 1-4 Weeks |
Direct exam by light microscopy of stained and unstained preparations. Culture on Sabouraud-
Dextrose and Mycocel agar plates. Identification by microscopy and characteristic growth on trichophyton agar. |
Fourty clinical and reference samples were processed for the identification of fungi in 2003. |
|
PARASITOLOGY |
|
| Parasitic Infections: Cryptosporidium, Giardia, Cyclospora, microsporidia, amoeba, roundworms, tapeworms, etc. Form Number
H-429 989 R3/97 |
Stool specimens only |
Ova & Parasite Kit contains: Formalin & PVA vials, with small plastic spoons attached to inside of lids, a stool collection container, 2 tongue depressers, lab requisition and instruction sheet.
Transportation: Stool specimens should be kept refrigerated before and during transport. Specimens in formalin and PVA can be transported without refrigeration.
Turnaround Time: 1-2 Days |
Specimens are concentrated using a formalin-ethyl acetate-extract procedure and examined by light microscopy using stained and unstained wet preparations.
Direct immunofluorescent microscopy is used for the simultaneous detection of Cryptosporidium oocysts and Giardia cysts. |
Fifteen different types of parasites were identified. Dual or multiple infestations were common. The most frequently detected parasite was: Giardia lamblia
|
| Pinworm Form Number
H-429 989 R3/97 |
Peri-anal |
Cellulose tape slide prep, applicator stick, swab, lab slip, instruction sheet. (transparent scotch tape, cellophane)
Transportation: Ambient
Turnaround Time: Same Day - 2 Days |
Microscopic examination of cellulose tape slide. |
One case of pinworm (Enterobius vermicularis) was detected in 2003. |
| Organism/ Infection | Specimen/ Site | Kit(s) Available at MHD - Laboratory (Specimen Handling) | Procedure | Statistics (2003) |
|---|
| |
CHLAMYDIA |
|
| Chlamydia trachomatis Culture Form Number
H-445 9/89-R10/98 |
Genital swabs: endocervical or Urethral, Newborn
Nasopharyngeal or eye swabs |
Need to use transport system specific for Chlamydia. Transport swabs or medium available upon request. Please call the lab.
Transportation: Refrigerate specimen soon after collection. DO NOT FREEZE!
Turnaround Time: 3-5 days |
Specimen inoculated into McCoy cells, incubated for 3 days, then stained with iodine for detection of Chlamydia spp. inclusion bodies.
Set-up twice weekly: Tuesday and Friday |
Approximately 300 in 2003. Positivity rate 12-15% annually.
Annual Chlamydia Testing: Milw. Health Dept. STD Clinic (1979-2003) |
Chlamydia trachomatis,
N.A. Amplification Form Number
H-445 9/89-R10/98 |
Male: urethral or urine
Female: endocervical or urine |
Need to use specific transport system for Becton Dickinson (BD Probe Tech)TM. Transport tubes available upon request. Please call the laboratory.
Transportation: Transport to the lab at room temperature.
Turnaround Time: 24 hours |
Utilizes the BD Probe TechTM nucleic acid technique to detect the presence of Chlamydia trachomatis.
|
Started use in 2003 with approximately 8000 specimens.
Positivity rate 18%. |
| Chlamydia pneumoniae, Culture Form Number:
H-445 9/89-R10/98 |
Nasopharyngeal or throat swabs, B.A.L., bronchial washings |
Washings; lavages; swabs without antibiotics that are designed for collection of Chlamydia. Transport media available upon request. Please call the lab.
Transportation: Refrigerate specimen soon after collection. Keep refrigerated during transport. DO NOT FREEZE!
Turnaround Time: 3-10 days |
Cultured in HEp-2 cells and incubated for 3 days. Cells then stained with fluorescent antibody to detect the presence of Chlamydia elementary bodies. All initial negatives are blind passaged.
Set up twice weekly. Tuesday and Friday. |
Organism reported to be responsible for 10% of all cases of pneumonia.
Causes illness in all age groups, but most severe disease in young children and the elderly. Prevalence increases with age. |
| |
VIROLOGY |
|
| Viral Culture: "Most common" human viruses will grow in the cells for culture. Form Number
H-445 9/89-R10/98 |
Collect specimen from appropriate site, i.e., throat for respiratory, rectal or stool swab for diarrhea, CSF for CNS disease. Lesion swabs for Herpes simplex or Varicella-zoster virus. |
Test tube of liquid MEM or viral culturette. Transport media available upon request. Please call the laboratory.
Storage: Immediately refrigerate or place on wet ice. May be held for up to 72 hours after collection. DO NOT FREEZE!
Transportation: Keep refrigerated or on wet ice during transport.
Turnaround Time: Typically, 1-10 days. Up to 21 days for slow growers. If necessary, phone for preliminary results. |
Specimens are inoculated onto at least six cell types: primary monkey kidney, HEp-2, human foreskin diploid, (HFS), human embryonic lung diploid, Madden-Darby canine kidney (also RD or BGMK cells for other seasonal viruses). Cells lines read daily for characteristic cytopathic effect (CPE) and all positives reported the day cell cultures become positive. Negatives reported after 9-11 days. For CMV, shell vials of human foreskin cells are incubated for 48 hours and then stained with fluorescent antibody for the presence of CMV- specific immediate-early antigen. In addition, HFS cell cultures are inoculated and held for 21 days, due to the slow growing nature of the virus. |
Approximately 2000 specimens are processed per year. Annual isolation rate is 15-20%. Most virus cultures become positive at 3 to 7 days - slightly faster or Herpes simplex viruses.
Influenza: 1967-2003 Enterovirus Monthly Composites 1988-2003 Rhinovirus Monthly Composites 1988-2003 RSV Monthly
Composites 1988-2003 |
| Herpes Simplex virus Form Number
H-445 9/89-R10/98 |
Oral/Genital swabs |
Viral transport medium or swab culturette appropriate for viral transport.
Storage: Keep swabs moist and refrigerated immediately after collection.
DO NOT FREEZE!
Transportation: Keep refrigerated and transport ASAP after collection.
Turnaround Time: 1-10 days. If necessary phone for preliminary results. |
Specimen inoculated onto 6 different cell types, as above. Cell lines read daily for characteristic CPE. Of those cultured that are positive for HSV-2, 30% are positive at 24 hours; 60% at 48 hours and 90% are positive at 72 hours. |
Annually, approximately 35% of genital specimens submitted for virus culture are positive for HSV.
For genital isolates, 85% are HSV-2 and 15% are HSV-1. |
| Rotavirus Antigen Detection Form Number
H-445 9/89-R10/98 |
Stool |
Collect fresh, raw stool in clean stool collection cup or swab.
Storage: Refrigerate, DO NOT FREEZE!
Transportation: Keep refrigerated
Turnaround Time: 1-2 days |
Enzyme Immunoassay (EIA-run daily) |
Annual winter epidemics from late December through March. Most infected are less than 3 years of age.
Rotavirus Monthly Composites 1988-2003 |
| Virus Serology Form Number
H-445 9/89-R10/98 |
Serum; Acute and Convalescent collected 10-14 days apart.
Acute and convalescent serums should be submitted together. |
Collect (1) 8 to 10 ml of whole blood without anticoagulant (red-top tube). Allow to clot. Keep refrigerated or (2) 4-5 ml of serum in serum separator tube.
Storage: Keep whole blood refrigerated. DO NOT FREEZE! Serum may be refrigerated for several days at 2-4oC. For extended storage, freezing at -20oC is recommended.
Turnaround Time: 1-7 days. |
The complement fixation (CF) test measures quantitative total antibody and is run once per week. Individual viral antigens are available, as well as antigen "panels" for respiratory, CNS, and exanthem (rash) viruses.
A four-fold increase in antibody titer is considered diagnostically significant.
See Test Listings for complete listings. |
Good test for respiratory virual infections, Mycoplasma pneumoniae and psittacosis. Also useful in the diagnosis of Herpes encephalitis, as well as for enteroviruses during the summer months. |
| Immune Status Testing for Measles, Mumps, Rubella, and Varicella Form Number
H-445 9/89-R10/98 |
Serum or whole blood in separator tube. |
As above.
Turnaround Time: 1-4 days |
Enzyme Immunoassay (EIA) for Measles, Mumps and Rubella IgG antibody.
Indirect Fluorescent antibody (IFA) for Varicella IgG antibody. |
The presence of IgG antibody is generally considered evidence of protective immunity under most circumstances. |
Specialty Serology:
M. pneumoniae IgM Parvovirus B19, Measles IgM Form Number
H-445 9/89-R10/98 |
Serum or whole blood in separator tube |
As above.
Turnaround Time: 1-4 days |
Enzyme Immunoassay (EIA) for M. pneumoniae IgM
Indirect Fluorescent antibody (IFA) for Parvovirus B19 IgG/IgM and Measles IgM
See Test Listings for complete
listings. |
The presence of IgM antibody is generally considered evidence of a recent infection with the virus or organism.
|
| Organism/ Infection | Specimen/ Site | Kit(s) Available at MHD - Laboratory (Specimen Handling) | Procedure | Statistics (2003) |
|---|
| |
CHEMISTRY |
|
Lead, Blood Form Number:
NB695 R6/03 |
Whole Blood,
venous or capillary |
Collect whole blood in purple top blood collection tube or Multivette brand collection tube containing EDTA or heparin. Pediatric minimum specimen is 250 microliters.
Storage: Refrigerate after collection
Transportation: Self-contained ziplock biohazard bag. Keep refrigerated
Turnaround Time: 24-48 hours |
Graphite Furnace Atomic Absorption Spectrometry |
Primarily used for pediatric lead poisoning detection.
Acceptable Ranges:
Children: 0-9 ug/100mL
Adults: 0-40 ug/100mL
Please see the Milwaukee Health Department recommendations for the interpretation of lead levels. |
| Hemocrit |
Whole Blood,
venous or capillary |
Equipment venous - purple top collection tube with heparin. Capillary micro hematocrit tube heparinized. Two ends to be sealed.
Storage: Refrigerate after collection
Transportation: In screwtop container inside self-contained ziplock biohazard bag. |
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| For information on incidence of Lead Poisoning in the City of Milwaukee call 414-225-LEAD or visit them on the Lead Poisoning web page. |