Wyoming Department of Health

Preventive Health and Safety Division

Public Health Laboratory

diag

DIAGNOSTIC SEROLOGY

Serologist Ruth Hagen (307) 777-6063
Microbiologist John Harrison (307) 777-6062

Routine testing is available in the laboratrories as descibed below. For more details see the WPHL Provider Manual or talk with one of the Laboratory Scientist listed above.

General Specimen Requirements (See also specific tests for additional requirements)
1. At least 1ml serum in leak proof plastic tube for each specimen/test combination.
2. Label specimen with patient name or unique identifier.
3. Sumit specimen with corresponding, completed, laboratory test request form.
4. If paired sera are sent, the acute specimen should be drawn within 3 days of the onset and the convalescent should be drawn 3 weeks after the onset.
5. For testing done by CDC or another State Health Lab it will be necessary to complete an additional test request form which includes information on date of onset, symptoms and immunization history. We will contact the submitter if this is necessary.
6. DO NOT HEAT INACTIVATE SAMPLES.

General Criteria for Rejection:
1. Gross hemolysis, lipemia, contamination
2. Improper Sample
3. No Name or identifier on specimen
4.Quantity not sufficient (QNS) - Please send sufficient volume if multiple tests are ordered.
5. Specimens not submitted by an approved health care provider or clinic

Packaging and Shipping of Diagnostic Specimens:
1. Ship body fluids and other human biological specimens as diagnostic specimens
2. Use mailers provided by WPHL or comperable approved shipping containers
3. Sender must follow postal or carrier shipping regulations associated with packaging & shipping

SEROLOGY

RPR Qualitative and Quantitative
VDRL
FTA-ABS
Rubella (German Measles)
Mumps
Varicella
Rubeola (Hard Measles)
Rocky Mountain Spotted Fever(RMSF)
Colorado Tick Fever
Lyme Disease
Miscellaneous Serologies

MICROBIOLOGY

Hantavirus Testing

RPR Qualitative and Quantitative
The RPR (rapid plasma reagin) test is a non-treponemal flocculation test used to screen for syphilis. Qualitative analysis is the initial screening of the undiluted specimen. Any specimen that is reactive initially will be quantitated. Quantitation is done by titering the specimen at doubling dilutions. Quantitation will establish a baseline from which changes in titer can be determined. Specimens that are reactive will be confirmed using FTA-ABS. Information on clinical presentation and evaluation can be obtained at the following website:http://www.cdc.gov/std

Reporting/Interpretation
1. Specimens will be reported as reactive or non-reactive.
2. Reactive specimens will also include the titer. The titer is the highest dilution giving a positive reaction.

Normal value
Non-reactive.

NOTES
1. Biological false positive reactions have been associated with some infectious diseases, narcotic addiction, and autoimmune diseases.
2. Testing of cord blood is not recommended.
3. Diagnosis of syphilis should not be made on a single reactive result without the support of a positive history or clinical evidence.

VDRL
The VDRL test is a non-treponemal serologic test for syphilis. It is a qualitative screening test that is recognized as a diagnostic test for neuro-syphilis. This test is performed only on cerebrospinal fluid (CSF).

Specimen Requirements
1. Only CSF may be submitted for analysis.

Reporting/Interpretation
1. Specimens will be reported as reactive or non-reactive.

Normal value
Non-reactive.

FTA-ABS
The FTA-ABS is a highly sensitive and specific confirmatory test for syphilis. All specimens with a reactive RPR will be tested. Specimens that come with requests for FTA-ABS, but are non-reactive by RPR will not be tested without consultation and approval of the program manager. Once a patient becomes FTA-ABS reactive, they may remain reactive for years, even if the patient has been treated properly.

Reporting/Interpretation
1. Specimens will be reported as reactive, non-reactive, or equivocal.
2. Fluorescence is graded on a 1+ to 4+ basis. If a specimen is 2+ or greater it is reported as reactive. If a specimen is < 1+, it is reported as non-reactive. If a specimen is graded as 1+, it will be reported as Equivocal, and another sample should be submitted for testing.

Normal value
Non-reactive.

Darkfield microscopy is not readily available at the Public Health Laboratory. Information concerning the availability of darkfield microscopy may be obtained by contacting the STD Program at (307) 777-7953.

Rubella (German Measles)
Qualitative screening for rubella antibodies is done by an enzyme linked fluorescent antibody technique. Detectable antibodies indicate past exposure to rubella either by disease or immunization. The Wyoming premarital law requiring that all females of childbearing years be tested for rubella and the Rh blood factor was repealed 22 Feb 95.

Reporting/Interpretation
1. Samples are screened undiluted.
2. Results will be reported as positive or negative.
3. Specimens that are reported as positive have detectable antibody, and the patient is considered to be immune if the assay is for screening purposes only.
4. Specimens that are reported as negative show no detectable antibody, and patients are considered to be non-immune or susceptible.

Normal value
Positive

Mumps
Specimens are routinely screened for mumps antibodies (IgG) using enzyme linked fluorescent antibody assay. This method can determine immune status on a single specimen. For symptomatic illness, an acute specimen may be submitted and will be referred to a reference laboratory for IgM testing. Please indicate "acute illness" on the requisition form. If an acute specimen is being submitted, please include the following:
1. Date of onset
2. Clinical symptoms
3. Immunization history

Reporting/Interpretation
Specimens that are sent in for routine screening will be reported as positive, negative, or equivocal. Positive results indicate evidence of antibody to mumps virus or vaccine and should be indicative of an immune status. Negative results show no evidence of antibody to mumps virus or vaccine and indicate that the individual is susceptible to mumps. Equivocal results are borderline and another specimen should be submitted since it is not possible to determine the immune status from this specimen.

FAQ Mumps

Mumps Key Facts

North Dakota Mumps Protocol

Varicella
Specimens are routinely screened for varicella antibodies (IgG) using enzyme linked fluorescent antibody assay. This method can determine immune status on a single specimen. For symptomatic illness, an acute specimen may be submitted and will be referred to a reference laboratory for IgM testing. Please indicate "acute illness" on the requisition form. If an acute specimen is being submitted, please include the following:
1. Date of onset
2. Clinical symptoms
3. Immunization history

Varicella Reporting/Interpretation
Specimens that are sent in for routine screening will be reported as positive, negative, or equivocal. Positive results indicate evidence of antibody to varicella virus or vaccine and should be indicative of an immune status. Negative results show no evidence of antibody to varicella virus or vaccine and indicate that the individual is susceptible to varicella. Equivocal results are borderline and another specimen should be submitted since it is not possible to determine the immune status from this specimen.

Rubeola(Hard Measles)
Specimens are routinely screened for rubeola antibodies (IgG) using enzyme linked fluorescent antibody assay. This method can determine immune status on a single specimen. For rash related illnesses, an acute specimen will tested by an IgM capture enzyme immunoassay. Please indicate "rash illness" on the request form. If an acute specimen is being submitted, please include the following:
1. Date of onset
2. Clinical symptoms
3. Immunization history

Rubeola(Hard Measles) Reporting/Interpretation
Specimens that are sent in for routine screening are reported out as positive, negative or equivocal. Positive results show evidence of antibody to measles virus, either wild virus or vaccination and should be indicative of an immune status. Negative results show no evidence of detectable antibody to rubeola virus and indicate that the individual is susceptible to rubeola. Equivocal results are borderline and another specimen should be submitted since it is not possible to determine the immune status from this specimen.

Rocky Mountain Spotted Fever (RMSF)
Indirect fluorescent antibody (IFA) testing is available for RMSF. Paired sera are preferred but a single sample can be tested. This procedure is used to demonstrate a four-fold rise in titer between acute and convalescent specimens. This would indicate a recent infection.

Rocky Mountain Spotted Fever (RMSF) Reporting/Interpretation
Results will be reported as the highest titer showing fluorescence. Reference reports with interpretations are provided.

Colorado Tick Fever
Lyme Disease
Miscellaneous Serologies

*Testing for Colorado Tick Fever (CTF) and Lyme Disease is available through CDC. Specimens for CTF need to be accompanied by a serology form and a completed CDC form #50.34.

*Specimens for Lyme Disease need to be accompanied by a serology form, a CDC history form #50.34, and a Lyme Disease report form. Paired sera are preferred but a single acute specimen will be accepted and tested for IgM.

*Specimens that are not accompanied by the appropriate forms WILL NOT be processed until these forms are completed.

The Public Health Laboratory works in conjunction with the Centers for Disease Control to offer a wide variety of immunological testing. These are handled on a case by case basis after conferring with the laboratory supervisor or director. All specimens to be forwarded to CDC must be accompanied by CDC history form #50.34. Processing of these specimens will not take place until after this form is received and properly completed. Under certain circumstances, an additional history form must be completed to aid in epidemiological study of the disease. Prior consultation is required if you have not previously sent in specimens of this nature. All bacterial, viral, mycological or rickettsial requests that are not specifically listed in this manual will be referred to another laboratory.

Hantavirus Testing
The recent outbreaks of Hanta Pulmonary Syndrome caused by the Sin Nombre virus (formerly Muerto Canyon) have prompted us to establish a serological test for IgG and IgM. Through training and reagents provided by the CDC, patient sera can be screened for Hantavirus. Clinical history consistent with Hanta Pulmonary Syndrome must be evident prior to submission of sample.

Hantavirus Testing Specimen Requirements
A minimum of 1 ml serum should be collected at the time the patient is admitted. A convalescent specimen should be collected 21 days after the first specimen. Fill out and submit a serology requisition form with each specimen.

Hantavirus Testing Shipment
The specimen should be shipped as soon as possible after collection. The tube should have the patient ID and date drawn. Place the specimen in a leakproof container and ship on ice packs and ship as diagnostic specimen according to shipping regulations. Please notify the laboratory of Hantavirus Specimens prior to shipment.

Hantavirus Testing Criteria for rejection
Improper sample.

Hantavirus Testing Reporting/Interpretation
This procedure is a CDC product and is not FDA approved for diagnostic testing. These results are subject to some interpretation. If the patient had an illness compatible with Hanta-associated respiratory illness and the specimen was drawn three or more weeks from onset of illness, a positive IgM is reasonable evidence that the illness was associated with a Hantavirus infection. A negative IgM response in a single specimen may not rule out the possibility of infection, as the specimen may have been collected too soon. A significant rise in IgG antibodies between acute and convalescent specimens is also suggestive of a recent infection. A positive IgG response in the absence of an IgM response may be indicative of a past infection. Results will be noted as "positive", "negative", or "equivocal", with a laboratory interpretation.

Hantavirus Testing Normal value
Negative.