Skip Maine state header navigation
Skip First Level Navigation | Skip All Navigation
 |
| Home | Contact Us |
|
Home
> HETL Sections > Microbiological Services
DIRECTORY
Adopted from the AMERICAN PUBLIC HEALTH ASSOCIATION (APHA)
HEALTH AND ENVIRONMENTAL TESTING LABORATORY DIRECTORY OF LABORATORY SERVICES MICROBIOLOGICAL SECTIONS HEALTH AND ENVIRONMENTAL TESTING LABORATORY TELEPHONE: (207) 287-2727 8:00 A.M. - 5:00 P.M. LABORATORY
Updated: September 1998 INTRODUCTION "The science and the art of preventing disease and prolonging life, and promoting physical and mental health and efficiency, through organized community efforts for the sanitation of the environment, the control of community infection, the education of the individual in principles of personal hygiene, the organization of medical and nursing service for the early diagnosis and preventive treatment of disease, and the development of the social machinery which will ensure to every individual in the community a standard of living adequate for the maintenance of health." C.-E. A. Winslow defined public health in this way in his publication, The Untilled Fields of Public Health, published in 1920. Despite all the changes in our nation and its health since that time, the definition seems remarkably apt. Although many infectious diseases are not the killers they were in Winslow's time, they are with us today, and without the constant vigilance of the public health system, could again overwhelm us. Surveillance for infectious and other diseases remains a cornerstone of public health, highlighted most recently by the Director of the U.S. Centers for Disease Control. The Laboratory plays a critical role as a part of the infrastructure of the surveillance system.The Health and Environmental Testing Laboratory, in the Department of Human Services, Bureau of Health, provides a wide range of chemical, microbiological, and radiological testing services to citizens and public and private agencies in the State of Maine, in order to protect the public's health from many acute and chronic health effects. These services, in the interest of the public, are provided at nominal cost to all Mainers. Fees, initiated in 1975, now provide about 80% of the Public Health Laboratory's operating budget, with 17% coming from direct State General Fund support, and 3% from federal sources.The Laboratory currently employs 59 people in administrative, technical, and support roles. It has an annual operating budget in excess of $2.5 million. The Health and Environmental Testing Laboratory is certified by the Health Care Finance Administration under the Clinical Laboratory Improvement Act of 1988 . The environmental testing services are certified by the US Environmental Protection Agency. This guide to laboratory services describes the clinical microbiology services offered to the public through the medical care system and through various Bureau of Health programs for disease surveillance and control. Other services, such as water testing and environmental analyses are described in this publication. TABLE OF CONTENTS COMPREHENSIVE LISTING OF MICROBIOLOGICAL TESTS PERFORMED INDIVIDUAL MICROBIOLOGICAL TESTS INCLUDING: BACTERIOLOGY, PARASITOLOGY, MYCOBACTERIOLOGY, MYCOLOGY, AND VIROLOGY ( LISTED ALPHABETICALLY ) (ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
(ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
(ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
(ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
(ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
(ALL ABBREVIATIONS ARE LISTED AT END OF TABLE)
ABBREVIATIONS USED IN TABLE: EIA = Enzyme Linked Immunoassay SPECIMEN COLLECTION AND HANDLING PROCEDURES VIROLOGY SECTION The Virology section currently offers a variety of Laboratory services not routinely available in the average hospital microbiology laboratory for assisting in the diagnosis in patients suffering from acute viral or nonviral infections. The section uses the three general methods for the detection and identification of viral (sometimes nonviral) infections: 1)Processing clinical specimens (e.g. throat swabs, stool specimens) in order to detect and identify viruses by cell culture; 2)Detection of IgG and or IgM antibody in single serum for diagnosis or demonstrating a significant four-fold or greater rise in antibody titer in acute and convalescent-phase sera to a known agent; 3)Out of State Premarital testing and diagnosis of Syphilis on Serum and Cerebrospinal Fluid. 4)Detecting microbial antigens in clinical specimens by immunofluorescence using specific conjugates to the agents. This last procedure is the most rapid but is routinely used only for a few infections at the present time; rabies (in another section), herpes encephalitis. Immune screening is offered for antibodies to agents capable of infecting the fetus in pregnant women including screening for cytomegalovirus, rubella and toxoplasmosis. In addition, immune screening is performed for rubella, rubeola, Varicella zoster (chickenpox), and Hepatitis B. Virus Isolation Virus isolation involves the testing of clinical specimens from patients (throat swabs, rectal swabs, vesicle fluid, cerebrospinal fluid, etc.) which are processed, treated with antibiotics and then inoculated into a suitable cell culture (cynomolgus monkey kidney or diploid human lung fibroblasts). The inoculated cell cultures are observed periodically for evidence of cytopathic effects for a period of six days (Herpes simplex only), 10 days (other suspect viruses), or 21 days (cytomegalovirus and Varicella zoster), before a report is sent. Cultures which exhibit evidence of viral growth are identified as specifically as possible with existing reagents. Collection of specimens for culture The collection of clinical material at the proper time, from the proper site and its handling will influence the recovery rate when specimens are received at the Laboratory. The following recommendations have been proposed for optimum recovery of the more labile viruses: 1)Specimens should be collected promptly, usually within the first 3 days and no longer than 7 days after onset of illness; 2)Almost any type of commercial swab can be used for most specimens (some laboratories have reported that calcium alginate fiber tips have inactivated herpes simplex virus and probably should not be used when another swab is available, (e.g. cotton or dacron tip swab). 3)Dry swabs used to collect clinical material should be immersed in a small volume of transport medium (1-2 ml) such as tryptosephosphate or nutrient broth. Commercial swabs containing medium for collection of virus specimens are also acceptable. Media containing charcoal or animal sera are not recommended, since the former ingredient may affect the rate of virus recovery and the latter may contain other viruses or filterable microorganisms. 4)The test specimen will yield more recoverable virus if it is stored at refrigerator temperature (2-8o C). If necessary, the specimen can be kept for up to several days prior to shipment. The infectivity of many viruses is rapidly lost by freezing specimens at -20o C and this procedure for storage is not recommended prior to shipment. 5)The specimen should be shipped in a foam container with a frozen gel pack. DO NOT USE DRY ICE! The package should be sent at the earliest possible time following current regulations for the shipping of diagnostic material. Throat, Nasal and Nasopharyngeal Cultures Swabs of the throat, oral cavity and nasal passages have been reported to yield the widest variety of viruses when taken in the early stage of illness (no later that 5-7 days after onset). A dry cotton swab is used to collect throat secretions by rubbing the back of the throat and any area of inflammation, ulceration or exudation. The swab is then broken or cut into a leakproof tube containing a small volume of medium (e.g. tryptose-phosphate broth with 0.5% gelatin). Nasal cultures are obtained by taking a cotton tip applicator moistened with broth and gently rotating it in a circular fashion after inserting the tip about 3/4 of an inch into the nares and then placing the applicator in broth and storing at 2-8o C. Nasopharyngeal specimens are obtained by taking a steel wire applicator and gently passing it through the nose to the posterior nasopharynx, leaving it for a few seconds and then removing it. Swabs tend to yield a lower recovery rate of virus than washings or aspirates but are more convenient to collect. Vesicle fluids and skin scrapings Specimens of vesicle fluids and cellular material from the base of lesions are best collected during the first 3 days of eruption when the recovery rate is the highest. Vesicle fluid can be aspirated using a 26 or 27 gauge needle attached to a tuberculin syringe or with a capillary pipette. The collected fluid should be rinsed into a small volume of transport medium. Open lesions can be swabbed to obtain both fluid and cells from the lesion base. The swab is then broken off into a vial of transport medium to preserve viable virus in the sample. Eye swab Any exudate or pus present in the eye should be carefully removed with a sterile swab. A second swab moistened with medium or saline should be used to rub the affected conjunctiva. This swab should be broken off into a vial of transport medium Cervical swab As with the eye swab, the first swab is used to clean the cervix of mucus and is discarded. A second swab is inserted about 1/4 inch into the cervical canal and rotated. If any lesions are seen, these should be collected on a swab and then the swab should be broken off into a vial of transport medium. Stool and rectal swabs Small amounts (1-4 g) of stool can be collected and placed in a small leakproof container. No transport medium should be added. A rectal swab is obtained by inserting a dry swab 1-2 inches past the anal sphincter, rotating it and then removing it. The swab should be broken off into a vial of transport medium and refrigerated. Commercially purchased viral collection swabs are also acceptable. Be sure to break the ampule at the base. Cerebrospinal fluid for Culture At least 2 ml should be collected into a sterile leakproof tube or vial. At least 1 ml should be obtained from infants. The specimen should not be diluted and should be kept in the refrigerator (2-8 oC) until sent to the Laboratory. Urine Clean-voided specimens (3-5 ml) should be stored at refrigerator temperature in sterile, leak-proof containers. Urine specimens are most useful for the isolation of cytomegalovirus in newborns and the best recovery rates occur if the specimen is held at 2-8 oC and processed within a few hours after collection. Autopsy and biopsy specimens Fresh tissue should be collected and placed in separate labeled sterile containers. Autopsy specimens in 1-2 gram portions are usually sufficient. Acceptable tissues from suspected cases include brain, lung, heart, lymph node and kidney. 1-2 ml of transport media should be added. Collection of blood sera Blood specimens should be collected without anticoagulants or preservatives. Routinely, a volume of 5 ml from an adult or 3 ml from a pediatric patient is sufficient for obtaining enough sera for testing. The blood specimen is allowed to clot at room temperature and the clot is then permitted to retract for about one hour. The specimen is then centrifuged for 10-15 minutes (at 1000-1300 g's) to separate the serum from the blood clot and any loose erythrocytes (we provide blood collection tubes with a gel separator which will physically separate the blood clot from the serum after centrifugation). The serum can either be refrigerated at 4-8 oC or frozen for antibody testing. Hemolysis or lysis of erythrocytes, generally will render the serum specimen unsuitable for most serological tests. It is important not to freeze whole blood specimens as this will result in hemolysis. Some laboratories have recommended that the blood specimens not be centrifuged until the clot has been allowed to form and retract either at room temperature or in the refrigerator. It is best to physically separate the serum from the blood clot within 24 hours of collection. Collection of cerecrospiral fluid CSF for Syphilis or Cryptococcus is the same as for culture except send CSF on gel seperator tubes for VDRL. The gel seperator interferes with results. Also CSF contaminated with serum is unsatisfactory. Serological testing Blood sera are routinely tested for screening or diagnosis of the following agents:
In general, a serological diagnosis of a recent infection requires the demonstration of a four-fold or higher antibody rise in paired serum specimens taken from the same patient in the acute and convalescent-phase of illness. The acute-phase serum is drawn as early as possible after onset while the convalescent-phase serum should be taken 10-14 days later. In cryptococcosis, the diagnosis of this fungal infection causing meningitis is made through the detection of the antigen in serum or cerebrospinal fluid and not through the formation of serum antibodies.In infections such as rubeola and rubella the culture procedure is inferior to the serologic procedure and is not recommended for routine use. Sera for serologic testing (e.g. paired sera from the same patient) collected during the acute and convalescent phases of illnesses can be stored in the refrigerator and sent when the second serum specimen has been collected. SPECIMEN COLLECTION AND HANDLING PROCEDURES BACTERIOLOGY SECTION ANAEROBE It is recommended that a pure isolate of the unknown organism be submitted for identification. A pure isolate serves to prevent undue delay in the identification of the organism. Submit the organism in thioglycollate, chopped meat or other growth supporting media. Avoid sending plates. Clinical material will be accepted but is not recommended due to the poor survival rate of the bacteria. Isolation should be attempted by the submitter. Clostridium difficile and Clostridium botulinum toxin testing is not done at the Health and Environmental Testing Laboratory, but is forwarded to CDC. Isolation and identification of pure isolates from clinical material is provided. Please call for instructions prior to submitting these specimens or in the case of suspected botulism.BACTERIA IDENTIFICATIONSubmit a pure isolate of the unknown bacteria for identification. Please provide prior laboratory results, i.e. gram stain, growth requirements, organism suspected etc. This serves to insure proper handling by this Laboratory upon receipt of the unknown. Serological typing for Haemophilus influenzae and grouping for Neisseria meningitidis is performed. Submit isolates on appropriate growth supporting media in screw capped tubes. Avoid sending plates.BORDETELLA PERTUSSIS Nasopharyngeal swabs are the specimen of choice. Transport media, FA slides and swabs are provided by the Health and Environmental Testing Laboratory. * Obtain a nasopharyngeal swab * Prepare at least 2 smears by gently rolling the swab over the slide surface in the ringed areas. (Insure that the frosted side is facing up before making smear.) * Insert the swab into the culture tube of Regan-Lowe media, either cut the wire (even with the top of the tube) or fold the wire down into the tube.* Cap, label, and place in the metal can. * Label the slide, air dry. Gently heat fix and place into the slide holder. * Place the can and slide holder into the box provided, and mail There is an estimated 95 % likelihood of a positive isolation in the first week of disease, dropping to less than 50 % in the fourth week.CAMPYLOBACTERCollect a freshly passed stool, uncontaminated by water or urine. Pick a portion of the stool with the swab provided and place into the vial of media. Leaving the swab in the vial, carefully break the swab at cap level. Screw the cap on tightly. Fill in the lab slip completely and forward these in the container provided. Stools for isolation and identification are accepted if submitted in appropriate transport systems. Chocolate agar slants are appropriate medium for submitting pure isolates for identification.DIPHTHERIAPure isolates for identification are accepted. Clinical material submitted in a suitable transport system, i.e. culturette, is also acceptable. Please call prior to the submission of specimens.DNA PROBE TESTING FOR NEISSERIA GONORRHOEAE AND CHLAMYDIA TRACHOMATIS A. Unisex Swab Specimen Collection Instructions: 1. Endocervical swabs (using amplified DNA probe assay): a. Remove excess mucus from the cervical os and surrounding mucosa using the cleaning swab (white shaft in the package with red printing). Discard the swab.b. Insert the specimen collection swab (blue shaft swab in the package with green printing) into the endocervical canal.c. Rotate the swab for 10-30 seconds in the endocervical canal to ensure adequate sampling.d. Withdraw the swab carefully; avoid any contact with the vaginal mucosa.e. Immediately place the swab into the GEN-PROBE transport tube.f. Break the swab shaft at the scoreline.g. Re-cap the tube tightly. h. Store the specimens at 2-30 °C until shipped at this temperature to arrive at the laboratory within 60 days of collection. 2. Urethral swab specimens (using amplified DNA probe assay): a. Patient should not have urinated for at least 1 hour prior to sample collection.b. Insert the specimen collection swab (blue shaft swab in the package with green printing) 2-4 cm into the urethra using a rotating motion to facilitate insertion.c. Once inserted, rotate the swab gently using sufficient pressure to ensure the swab comes into contact with all urethral surfaces. Allow the swab to remain inserted for 2 to 3 seconds.d. Withdraw the swab.e. Immediately place the swab into the GEN-PROBE transport tube.f. Break the swab shaft at the scoreline.g. Re-cap the tube tightly. h. Store the specimens at 2-30° C until shipped at this temperature to arrive at the laboratory within 60 days of collection. 3. Urine specimen (using amplified DNA probe assay):
SALMONELLA Isolation from clinical, food or environmental specimens. Isolates should be submitted on agar slants, for identification and serotyping. Serotyping is currently done at no charge to the submitter.E.coli: 0157:H7Stools as well as pure isolates of E. coli are accepted for isolation, identification and serotyping. NEISSERIA GONORRHOEAEFA reference slides and cultures are accepted. If warranted, identification and beta lactamase testing are done from culture. Submit viable cultures on appropriate media in a CO2 atmosphere.PARASITIC DISEASESThis service is no longer routinely provided by the laboratory. Please call for assistance during outbreaks or in identifying unusual parasites. Intermittent passage of parasites by the host may require multiple specimens taken over a period of several days. Passed stool specimen gives better test results than those obtained by catharsis or sigmoidoscopy. Stool specimen obtained by enema may be unsatisfactory for examination if contaminated by barium, etc. Stools should not be submitted until one week after the use of barium. Certain medications may also interfere with the detection of intestinal protozoa. Examples are: mineral oil, bismuth, antidiarrheal preparations, and some antibiotics. Stools are examined for:
Recovered helminths should be placed in a preservative solution such as formalin for shipment to the Laboratory. Blood parasites are referred. Prior notification is necessary before submission of specimens to the Health and Environmental Testing Laboratory.
SPECIMEN COLLECTION AND HANDLING PROCEDURES RABIES
RECOMMENDATIONS FOR SUBMITTING ANIMALS FOR TESTING Animals to be tested should have bitten or exposed a person or a domestic animal. The laboratory recommends that you consult with one of the rabies staff in cases where questionable exposure may be a factor to determine whether the animal should be tested. A fact sheet on the rabies website is available. A rabies manual has been published for emergency personnel (physicians, veterinarians and animal control officers). The animal should be killed in a manner which would not damage brain tissue (i.e. do not shoot animal in head or club it to death). Whenever possible, only the head should be brought to the laboratory in a leakproof plastic bag. Skunks should be tightly sealed to minimize skunk odors when brought into the laboratory. Smaller animals (e.g. bats) can be brought in intact. If the animal cannot be brought in shortly after death, to minimize decomposition, it should be refrigerated (2-80°C) but not frozen; it can be stored up to several days in this manner. A clinical history, including the names of physicians or veterinarians consulted, must be included with the specimen if the specimen is sent to the Health and Environmental Testing Laboratory by bus or other means. If the specimen is hand carried, the history will be taken upon receipt. All results will be reported by telephone whenever possible. A written report of the laboratory results will also be sent. Routinely, specimens submitted on the morning of a work day will be tested and reported in the afternoon. Specimens received in the afternoon of a work day will be tested and reported on the next work day, unless there is an emergency involving the bite of a high-risk animal ( raccoon, skunk, fox, bat, woodchuck) to a human patient. SPECIMEN COLLECTION AND HANDLING PROCEDURES MYCOBACTERIOLOGY AND MYCOLOGY CONTAINERS
MYCOBACTERIOLOGY Specimens
Reporting: Acid Fast Bacilli (AFB) smears are prepared, read and the results are reported the same day. All specimens are also cultured the same day.
Identification: The Gen-Probe, DNA PROBE test is offered for a rapid identification of: M. tuberculosis complex, M. avium complex, and M. gordonae. This method requires growth that is no more than four weeks old. Those Mycobacteria not identified by Gen-Probe are identified by the routine biochemical tests which require up to three weeks after culture growth for completion.
MYCOLOGY SPECIMENS
If there are any questions regarding Mycobacteriology or Mycology, please call for additional information.SERVICES AVAILABLE AT THE CENTERS FOR DISEASE CONTROL (CDC) Occasionally, there is a need for specialized laboratory testing for rare or exotic diseases which require the resources of the Center for Infectious Diseases, Centers for Disease Control. Because of the large number of tests for various microbial infections, inquiries for consultative or testing services should be made to the appropriate microbiology section within the Health and Environmental Testing Laboratory (e.g. Virology, Bacteriology, or Mycobacteriology) if culture material is being sent. All questions regarding serological testing should be directed to the Virology section. The general requirements for serum specimens are: 1. CDC form 50.34 MUST be completed as fully as possible in order to provide an accurate clinical history of the patient (e.g. Date of rash onset for measles specimens is important in determining suitability of the specimen for the test ordered and if any other test or sample type would be more appropriate). Failure to comply with this can result in the rejection of the serum specimen by the CDC. 2. Serum specimens should be aseptically collected to minimize contamination. The usual volume of blood drawn from adults is 8-10 ml although 3-4 ml from pediatric patients may be sufficient. The blood specimen is allowed to clot at room temperature for about one hour and then centrifuged to collect the serum into a sterile tube. 3. Paired serum specimens from the same patient are preferred and in some cases, required by the CDC (exceptions are tests for parasitology, mycology and syphilis). The acute-phase serum should be obtained as soon as possible after clinical onset while the second, convalescent-phase serum should be taken 2-4 weeks later depending upon the suspected disease. 4. Human Parvovirus testing is available, but because of the ubiquitous nature of these viruses, primary diagnosis by serologic testing or virus isolation cannot be routinely offered. These services may be provided by prior consultation and arrangement with the Division of Viral and Rickettsial Diseases for outbreaks or cases of unusual public health significance. (e.g. Patients with suspected unusual manifestation of B-19 infection requires prior approval from Dr.William Gary at the CDC for each patient). 5. Occasionally a bacterial isolate cannot be identified by the Bacteriology section of the Health and Environmental Testing Laboratory and will be referred to the CDC for identification. 6. When Clostridium botulinum is suspected, please call the Bacteriology Section for information and instructions on shipment and handling. Contact with the CDC is necessary prior to shipping material for analysis. 7. Toxin testing for E. coli 0157:H7 is done by the CDC. Isolation, identification and serotyping is done by the Health and Environmental Testing Laboratory. Specimens are no longer being routinely sent for toxin testing. 8. Staphylococcal phage typing and toxin testing require prior approval from the CDC. Usually single cases are not accepted unless epidemiologically significant.
NOTIFIABLE DISEASES NOTIFIABLE CONDITIONS LIST MAINE DEPARTMENT OF HEALTH AND HUMAN SERVICES, BUREAU OF HEALTH
Who must report:
What to report:
Disease reports must include as much of he following as is known:
HOW TO REPORT:
Complete Rules for the Control of Notifiable Conditions at http://www.maine.gov/dhs/ddc/DiseaseReporting.htm
The Laboratory Certification and Improvement Program provides the following services: Evaluates and certifies drinking water/environmental Evaluates independent clinical laboratories for State Evaluates Substance of Abuse testing laboratories Evaluates and issues permits to Health Screening Sponsors, coordinates and conducts educational Monitors proficiency testing performance for
The Department will consider reducing the fee for certain tests for Public Agencies, if requested. Public Agencies are governmental units and non-profit health agencies receiving financial support from the Department of Human Services for public testing or services.
NON-DISCRIMINATION NOTICE: In accordance with Title VI of the Civil Rights Act of 1964 (42 U.S.C. 2000d et seq.), Section 504 of the Rehabilitation Act of 1973, as amended (29 S.S.C. 794), the Age Discrimination Act of 1975, as amended ( 42 R.S.C. 6101) et seq.), and Title IX of the Education Amendments of 1972, the Maine Department of Health and Human Services does not discriminate on the basis of sex, race, color, national origin, handicap, or age in admission or access to or treatment or employment in its programs or activities. Ann Twombly, Affirmative Action Officer, has been designated to coordinate our efforts to comply with the U.S. Department of Health and Human Services regulations (45 C.F.R. Parts 80, 84, and 91) and the U.S., Department of Education (34 C.F.R. Part 106) implementing these Federal laws. Inquiries concerning the application of these regulations and our grievance procedures for resolution of complaints alleging discrimination may be referred to Ann Twombly at 221 State Street, Augusta, Maine, 04333. Telephone: (207) 287-3488 (Voice) or 1-800-332-1003 (TTY), or to the Assistant Secretary of the Office of Civil Rights, Washington, D.C.. |
| Copyright © 2005 All rights reserved. |
