Delaware Public Health Laboratory Specimen Collection Procedures for Identification of Bioterrorism Organisms DISEASE/AGENT - Anthrax Bacillus anthracis SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical *Vesicular Stage: use Culture: rule-out and Grown isolate on agar 24-72 sterile swaps to culture confirmation plate sent at room hrs collect fluid from available on grown temp intact vesicles. The organism. organism is best demonstrated in this stage. *Eschar stage: without PCR: available on Grown isolate on agar 4-8 PCR requires culture removing eschar,insert grown organism only. plate sent at room hrs confirmation swap beneath the edge temp of eschar, rotate and collect lesion material. Lesion biopsy: collect Histochemical stain: Double bag, sent at Will Contact DPHL to biopsy in sterile available through CDC room temp need to request CDC tube with 10% formalin. for high suspect cases be sent reference number and only to CDC transportation options May require Chain of Custody *Stool: collect 5-10g Culture: rule-out and Grown isolate on agar 48-72 Minimal recovery in a clean, sterile, culture confirmation plate sent at room hrs leak proof container. available on temp grown organism. PCR available on Grown isolate on agar 4-8 PCR requires culture grown organisms plate sent at room temp hrs confirmation *Blood Culture: collect Culture: not available Grown isolate on agar 24-72 Blood culture is positive per institution’s plate sent at room hrs in late stages of disease procedure for routine temp Blood culture cannot be blood culture. performed at DPHL Blood culture is incubated by facility PCR available on grown Grown isolate on agar 4-8 PCR requires culture drawing the culture organism only plate sent at room hrs confirmation and screened for temp growth of the organism. Blood: purple top tube Culture: not available May require Chain of Custody (EDTA), serum, plasma PCR: available on refrigerate 8-24 PCR requires culture original specimen at 2-8C hrs. confirmation Pleural fluid, Culture: rule-out and Grown isolate on agar 24-72 Sputum: minimal recovery transtracheal culture confirmation plate sent at room hrs aspirates, fresh available on grown temp tissue, frozen organism. tissue, sputum: collect PCR: available on Refrigerate at 8 -24 May require Chain of custody specimen in a sterile, original specimen 2-8C hrs PCR requires culture leakproof container. confirmation Environmental Powder, letter, etc.: Culture: available Dacron or polyester 24-72 Chain of Custody required collect at least two swabs or a culturette hrs non-cotton swabs, PCR: available Dacron or polyester 4-8 Chain of Custody required place in separate swabs no cuturettes hrs PCR requires culture sterile containers. confirmation Food Notification of DPHL Culture: available Refrigerated samples 2-5 Chain of Custody required by DPH Epidemiology PCR: available & those collected at days PCR requires culture Section is necessary Send all suspect room temp should be confirmation as soon as possible food in a sterile leak transported at 4C. Submitter must contact due to the large proof container. Transport frozen DPH Epidemiology Section amount of media Minimum needed to samples on dry ice. concerning any suspected necessary to process test: 25 grams food related infections. food. * Cultures from clinical specimens should be performed by the sentinel laboratory ____________________________________________________________________________________________________________________________ DISEASE/AGENT - Botulism - Toxins are produced by certain strains of Clostridium botulinum SPECIMEN SELECTION FROM CLINICAL VOLUME REQUIRED TEST COMMENTS SAMPLES TRANSPORT TEMP METHODS Specimen Type Clinical Syndrome A=Autopsy specimen may be used Mouse Bioassay will be Mouse Bioassay Anaerobic Culture: organism is isolated performed if less of is only and identified as Clostridium botulinum. Food Infant Wound Intentional than 10cc of extracted confirmatory Extraction of toxin from organism may borne Release * supernatant available test available tested by DIG-ELISA and Mouse Bioassay. Anaerobic Culture confirmatory test. Enema X X X X 20 cc Anaerobic Purge with a minimal amount of Fluid 4°C Culture Mouse sterile nonbacteriostatic water Bioassay to minimize dilution of toxin Gastric X, A 20 cc Anaerobic Collect up to 20 cc Fluid 4°C Culture DIG-ELISA Mouse Bioassay Intestinal A A 20 cc Anaerobic Autopsy: intestinal contents from Fluid 4°C Culture various areas of the small and Mouse Bioassay large intestines should be provided. Nasal X Anaerobic Swab Anaerobic For aerosolized botulinum toxin Swab Room Temp Culture exposure, obtain nasal culture for Mouse Bioassay C. botulinum and serum for mouse toxicity testing. Serum X, A X X 10-12 cc Mouse Bioassay Serum should be obtained as soon as 4°C possible after the onset of symptoms and before antitoxin is given. A minimum of 10 cc of serum (20cc of whole blood) is required for mouse toxicity testing. In infants, serum is generally not useful, since the toxin is quickly absorbed before the serum can be obtained. Stool X X X X 10-50 g Anaerobic Botulism has been confirmed in 4°C Culture infants with only “pea sized” DIG-ELISA stools. Please note: Mouse anticholinesterase given orally, Bioassay in patients with myasthenia gravis, has been shown to interfere with toxin testing. Vomitus X 20 cc Anaerobic Collect up to 20 cc 4°C Culture Mouse Bioassay Wound/Tissue X Anaerobic Swab Anaerobic Exudates, tissue, or swabs must be or Culture collected and transported in an Transport System Mouse anaerobic transport system. Room Temp Bioassay Samples from an enema or feces should also be submitted since the wound may not be the source of botulinum toxin. Environmental X X 3-6 swabs Anaerobic Environmental Swabs Sample Room temp Culture DIG-ELISA Chain of Custody required Mouse Bioassay Food X X X 25-50 g Anaerobic Chain of Custody required 4°C Culture DIG-ELISA:>5ml Mouse Bioassay Food that supports C. botulinum liquid or 25g DIG-ELISA growth will have a pH of 3.5-7.0. solid. Regrig 4C Most common pH is 5.5-6.5. Bioassay The most common pH is 5.5-6.5. Submit food in original container, placing individually in leak proof sealed transport devices. Submitter must contact Epidemiology Section of DPH concerning any suspected food related infections. *Anaerobic culture for suspected c. botulinum specimens is performed by LRN reference laboratory. **DPHL only performs DIG-ELISA testing on Environmental and Food samples for botulism. It will detect types A, B, E & F. All other specimens will be sent to reference laboratory ***Clinical specimens from an intentional release may require a Chain of Custody. _____________________________________________________________________________________________________________________________ DISEASE/AGENT - Brucellosis (Brucella melitensis, Brucella abortus, Brucella suis, Brucella canis) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical *Bone marrow, spleen, Culture*: rule-out and Grown isolate on 5-10 days *Cultures from or liver: collect culture confirmation agar plate sent at clinical specimens Acute, per institution’s available on grown room temp should be performed subacute, surgical/pathology organism. by the sentinel or chronic procedure. laboratory. PCR: available on Grown isolate on 4-8 hrs PCR requires grown organism only. agar plate sent at culture room temp confirmation *Blood Culture: Culture: rule-out and Grown isolate on 5-10 days Blood culture can collect per culture confirmation agar plate sent at not be peerformed institution’s available on grown room temp at DPHL procedure for routine organism. blood culture. Blood culture is PCR: available on Grown isolate on 4-8 hrs PCR requires incubated by facility grown organism only. agar plate sent at culture drawing the culture room temp confirmation and screened for growth of the organism. Blood: whole blood, Culture: not available --- --- --- at least 200 ul PCR available on Refrigerate 2-8C 4-8 hrs PCR requires grown organism only Shipping requires culture freezing -20C confirmation May require chain of Custody Serum: collect 10-12cc Serologic diagnosis: Refrigerate 2-8C Will need Contact DPHL to acute phase specimen 1. Single titer to be sent request CDC as soon as possible greater than 1:160 shipping to CDC reference number after disease onset. 2. 4-fold rise requires freezing and transportation Collect a convalescent 3. IgM positive at -20°C options specimen obtained 21 May require days later. Chain of Custody Environmental Powder, letter, etc.: Culture: available Use Dacron or 5-10 days Chain of Custody collect at least two polyester swabs or required non-cotton swabs, a culturette place in separate sterile containers. PCR: available on Use Dacron or 4-8 hrs for Chain of Custody original specimen polyester swabs, organism required and grown organisms no culturettes 8-24 hrs for PCR required environmental culture confirmation Food Notification of DPHL Culture and PCR: Refrigerated samples 5-10 days Chain of Custody by DPH Epidemiology available & those collected at required Section is necessary Send all suspect room temp should be PCR requires as soon as possible food double bagged in transported at 4C. culture due to the large in a leak proof Frozen samples should confirmation amount of media container. Minimum be transported on Submitter must necessary to process needed to test: 25g dry ice contact DPH food. Epidemiology Section concerning any suspected food related infections. ___________________________________________________________________________________________________________________________ DISEASE/AGENT - Glanders (Burkholderia mallei), Meliodidosis (Burkholderia pseudomallei) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical *Abscesses: collect Culture: rule-out Grown isolate on 4-7 days *Cultures from using a culturette and culture agar plate sent at clinical specimens confirmation room temp should be performed available on by the sentinel grown organism. laboratory if possible PCR: available on Grown isolate on 4-8 hrs PCR requires grown organism only agar plate sent at culture (see comments) room temp confirmation *Urine, tissue Culture: rule-out Grown isolate on 4-7 days *Cultures from aspirates: collect and culture agar plate sent at clinical specimens into a sterile, leak confirmation room temp should be performed proof container. available on grown by the sentinel organism. laboratory if possible PCR: available on Grown isolate on 4-8 hrs PCR requires grown organism only agar plate sent at culture (see comments) room temp confirmation *Blood Culture: Culture**: rule-out & Grown isolate on 4-7 days Blood culture is collect per culture confirmation agar plate sent at rarely positive institution’s available on grown room temp Blood culture can procedure for routine organism. not be performed blood culture. Blood at DPHL culture is incubated by facility drawing PCR: available on Grown isolate on 4-8 hrs PCR requires the culture and grown organism agar plate sent at culture screened for growth only (see comments) room temp confirmation of the organism. Blood: serum Culture: not available --- --- --- (at least 200 ul) PCR: available on Refrigerate at 8-24 hrs May require grown organism only 2-8C Chain of Custody (see comments) Transport on dry ice PCR requires Culture confirmation Environmental Powder, letter, etc.: Culture: available Use Dacron or 8-24 hrs Chain of custody collect at least two polyester swabs required non-cotton swabs, place in separate PCR: available on Use Dacron or 4-8 hrs for Chain of Custody sterile containers. original specimen polyester swabs organism required on and grown organisms 8-24 hrs for environmental environmental samples PCR requires culture confirmation Food No method available at DPHL or CDC *PCR detects both Burkholderis mallei and Burkholderia pseudomallei. Culture is necessary to differentiate between species. **Cultures from clinical specimens should be performed by the sentinel laboratory if possible. ____________________________________________________________________________________________________________________________ DISEASE/AGENT - Q fever (Coxiella burnetti) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical 2 Bloods in serum Indirect Refrigerate at 12-24 hours May require separator tube: Immunofluorescent 2-8C after initial Chain of Custody 1st tube: collect antibody (IFA) set up first tube as soon as test for detection possible within four of specific antibodies days of illness in serum 2nd tube: collect about two to three Standard PCR: --- --- --- weeks after onset no available of disease EDTA blood collected IFA: not available --- --- --- in acute phase before antibiotic therapy Real-Time PCR: Refrigerate at 24-48 hrs May require available 2-8°C Chain of Custody Affected tissues to IFA: not available --- --- --- include autopsy specimens or tissue Standard PCR: Refrigerate at 24-48 hrs May require biopsies, blood clots, available 2-8°C if lengthy Chain of Custody and cell cultures delay before (or cell culture reaching lab, freeze supernatants), collect at -70°C in sterile leak proof screw-capped containers Environmental Powder, letter, etc.: IFA: not available --- --- --- collect at least two non-cotton swabs, place Real-Time PCR: Do not use any 4-8 hrs for Chain of Custody in separate sterile available on transport media organism required original specimen Refrigerate at 8-24 hrs for 2-8C environmental PCR requires PCR requires culture confirmation Food No method available at DPHL or CDC _________________________________________________________________________________________________________________________ DISEASE/AGENT - Ricin toxin - Ricin biomarker determiatnion - Ricin toxin is detected using the TRF method SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical Urine: collect LC/MS/MS: available* Freeze at -20-80C 12-36 hrs. Method available minimum 25-50mL following IATA PI through LRN in plastic urine 650 & CDC Chemical network - specimen cup with Exposure Guidelines anticipated at O-ring DPHL Environmental *Swabs (cotton, Time-resolved Transport at room 8-24 hrs *Do not collect polyester, Dacron, fluorescence assay temperature samples in rayon, or foam) (TRF): available containers made of glass. If * Wipes (non-cotton possible avoid gauze, polyester collection from blend, e.g., “Handi- any glass surface Wipes”) Chain of Custody *Plant material in an required envelope, paper, powder, water, soil (2.0±0.1g) Food *Food, drink: collect Time-resolved Refrigerate 8-24 hrs *Do not collect 2.0±0.1g, place in fluorescence assay 2-8C samples in sterile, leak proof, (TRF): available containers made plastic container of glass. If possible avoid NO GLASS CONTAINERS collection from any glass surface Chain of Custody required __________________________________________________________________________________________________________________________ DISEASE/AGENT - Ricin communis - DNA from the Ricin communis plant is detected using PCR method SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical No method available at --- --- --- --- DPHL or CDC Environmental *Swabs (cotton, RT-PCR: available Transport at room 8-24 hrs *Do not collect polyester, Dacron, temperature samples in rayon, or foam) containers made of glass. If * Wipes (non-cotton possible avoid gauze, polyester collection from blend, e.g., “Handi- any glass surface Wipes”) Chain of Custody *Plant material in an required envelope, paper, powder, water, soil (2.0±0.1g) Food *Food, drink: collect RT-PCR: available 2-8°C 8-24 hrs *Do not collect 2.0±0.1g, place in samples in sterile, leak proof, containers made plastic container of glass. If possible avoid NO GLASS CONTAINERS collection from any glass surface Chain of Custody required ____________________________________________________________________________________________________________________________ DISEASE/AGENT - SEB - Staphylococcal Enterotoxin B SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical No method available at --- --- --- --- DPHL or CDC Environmental *Swabs (cotton, Time-resolved Place into a leak 8-24 hrs *Do not collect polyester, Dacron, fluorescence assay proof, sealed, samples in rayon, or foam) (TRF): available non-glass containers made transport device of glass. If *Wipes (non-cotton possible avoid gauze, polyester collection from blend, e.g., “Handi- any glass surface Wipes”) Chain of Custody *Plant material in an required envelope, paper, powder, water, soil (2.0±0.1g) in sterile (if possible), leak proof container. NO GLASS CONTAINERS Food *Food, drink: collect Time-resolved Place into a leak 8-24 hrs *Do not collect 2.0±0.1g, place in fluorescence assay proof, sealed, samples in sterile (if possible), (TRF): available non-glass containers made leak proof, plastic transport device. of glass. If container possible avoid collection from NO GLASS CONTAINERS any glass surface Chain of Custody required ___________________________________________________________________________________________________________________________ DISEASE/AGENT - Tularemia (Francisella tularensis) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical **Bronchial/tracheal Culture: rule-out Grown isolate on 24-72 hrs *Cultures from wash**: collect per and culture agar plate set at clinical specimens institution’s proc. confirmation room temp should be performed in area dedicated to available on by the sentinel collection of grown organism. laboratory if respiratory specimens possible under isolation/ containment PCR: available on Grown isolate on 4-8 hrs PCR requires circumstances, i.e., grown organism only agar plate set at culture isolation chamber/ room temp confirmation ”bubble” *Sputum/throat: Culture: rule-out Grown isolate on 24-72 hrs Minimal recovery-- collect routine throat and culture agar plate set at not recommended culture using a swab or confirmation room temp collect expectorated available on sputum into a sterile, grown organism. leak proof container. PCR: available on Grown isolate on 4-8 hrs PCR requires grown organism agar plate set at culture room temp confirmation *Blood Culture: collect Culture: rule-out Grown isolate on 24-72 hrs Blood culture is per institution’s and culture agar plate set at rarely positive procedure for routine confirmation room temp Blood culture blood culture. Blood available on can not be culture is incubated grown organism. performed at DPHL by facility drawing the culture and screened PCR: available on Grown isolate on 4-8 hrs PCR requires for growth of the grown organism agar plate set at culture organism. room temp confirmation **Ulcer/wounds/tissue: Culture: rule-out Grown isolate on 24-72 hrs --- collect 2 swabs using and culture agar plate set at a culturette, collect confirmation room temp tissue in sterile available on container. grown organism. PCR: available on Grown isolate on 4-8 hrs May require grown organism agar plate set at Chain of Custody room temp PCR requires culture confirmation Blood (EDTA): one Culture: not available --- --- --- purple top tube PCR: available on Refrigerate at 8-24 hrs May require original specimen 2-8C Chain of Custody PCR requires culture confirmation Environmental Powder, letter, etc.: Culture: available Use Dacron or 2-7 days Chain of Custody collect at least two polyestyer swabs or required non-cotton swabs, a culturette place in separate sterile containers. PCR: available on Use Dacron or 4-8 hrs for Chain of Custody original specimen polyester swabs, organism required on and grown organisms no culturettes 8-24 hrs for environmental environmental samples PCR requires culture confirmation Food Notification of DPHL Culture and PCR: Refrigerate samples 2-7 days Chain of Custody by DPH Epidemiology available & those collected at required Section is necessary Send all suspect room temp should be PCR requires as soon as possible food in a leak transported at 4C. culture due to the large proof container. Transport frozen confirmation amount of media Minimum needed samples on dry ice. Submitter must necessary to process to test: 25 grams contact DPH food. Epidemiology Section concerning any suspected food related infections. ___________________________________________________________________________________________________________________________ DISEASE/AGENT - Vaccinia virus "pox" type virus related to smallpox and is used to induce immunity against smallpox. SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical Dried vesicular fluid Virology tests: none on a slide available ----- ------ ------ PCR: available Slides must be 12 hrs. from Notes: These packed in a plastic receipt of results are slide container sample. preliminary. Refrig at 2-8C Positive PCR results must be confirmed by CDC Clinical speci. may require Chain of Custody. Vesicular tissue Virology tests: none ------- ------- ------- (Skin from roof available. of vesicle). PCR: available Place in sterile 12 hrs. from These results are leak proof, receipt of preliminary. unbreakable container samples Positive PCR results must be confirmed by CDC. __________________________________________________________________________________________________________________________ DISEASE/AGENT - Chicken pox virus (Varicella zoster virus VZV) - used as a rule-out test for low-moderate risk patients for possible smallpox SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical Vesicle fluid or Direct DFA: available Refrigerate at 12-24 hrs --- lesions: collect 2-8C vesicular fluid on a polyester swab, scabs from a Viral culture: Refrigerate at 4-10 days --- crusted lesion, appropriate cell line 2-8C skin from the top will be inoculated of a vesicle or at DPHL pustule PCR: available Do not use viral 8-12 hrs PCR requires transport media viral culture Transport at room confirmation temperature Blood: 10 cc of IFA: available Refrigerate at 12-24 hrs --- blood collected in 2-8C a serum separator tube Viral culture: --- --- --- not available PCR: not available --- --- --- Environmental No method available at --- --- --- --- DPHL or CDC Food No method available at --- --- --- --- DPHL or CDC ____________________________________________________________________________________________________________________ DISEASE/AGENT - Smallpox virus (Variola major) *NOTE: High risk patient testing (see other chart for low to moderate risk patient testing) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME ****Utilize CDC Guide D Precautions for collection of specimens from High-risk patients**** Clinical Fresh lesions: if CDC will determine Refrigerate at *See below *Specimens from possible, collect tests to be performed 2-8C patients in the For patients several Dacron or high-risk category in the high polyester swabs, for possible risk place each swab in smallpox will not categories separate viral be tested at DPHL of the rash transport media illness CDC requires protocol: Autopsy specimens: CDC will determine Ship frozen on dry ice. *See below BioSafety Level 4 Including portions tests to be performed facilities to be 1. A suspected of skin containing used for smallpox case of small lesions, liver, kidney, testing and pox should etc., for virus specimens will be reported isolation should be therefore be immediately to frozen referred to CDC DPH for review. for testing. Biopsy: Place dried CDC will determine Package in slide box 2. And if, vesicular fluid on tests to be performed Do not freeze Clinical Specimens after review, a slide ("touch may require a true case prep") Chain of Custody of smallpox is still Vesicles or pustules: CDC will determine Refrigerate at --- See above suspected, DPH use scapel to open, tests to be performed 2-8C will contact remove top of the CDC’s Poxvirus vesicle or pustule Section for and place the skin approval to of the vesicle top send specimens. into a 1.5-2.0 ml screw capped plastic 3. DPHL will vial. contact CDC for review Scab lesions: place CDC will determine Refrigerate at --- See above of the two scabs in each tests to be performed 2-8C packaging/ of two 1.5-2.0 ml shipping screw capped plastic requirements vial. and to seek coordination Blood: 10 cc of CDC will determine Refrigerate at --- See above of transport/ blood collected in tests to be performed 2-8C shipment of a serum separator the specimens tube to CDC. Environmental No method available at --- --- --- --- DPHL or CDC Food No method available at --- --- --- --- DPHL or CDC *CDC will notify DPHL of results. DPHL will then notify submitter as soon as possible. ****CDC Smallpox Draft Guide C, Part 1, Infection Control Measures for Healthcare and Community Settings, Guide F, Environmental Control f Smallpox Virus, http://www.bt.cdc.gov/agents/smallpox/infection-control/ ______________________________________________________________________________________________________________________ DISEASE/AGENT - Smallpox virus (Variola major) *NOTE: Low risk patient testing only (see other chart for high risk patient testing) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical Fresh lesions: if Cell culture Refrigerate at 4-10 days For patients in possible, collect 2-8C low and moderate For patients several Dacron or risk categories in the low polyester swabs, according to the and moderate place each swab in evaluation criteria risk categories separate viral in the CDC rash according to transport media illness protocol the evaluation DPHL can perform criteria in Autopsy specimens: Cell culture Ship frozen on dry 4-10 days rule-out tests THE CDC rash Including portions ice. for Smallpox which illness of skin containing includes vaccinia, protocol: lesions, liver, kidney, VZV (Varicella- etc., for virus zoster virus), and DPHL can perform isolation should be herpes simplex rule-out tests frozen. virus. for Smallpox which includes Biopsy: Place dried PCR Package in slide 12 hrs. from Clinical Specimens vaccinia, VZV vesicular fluid on box, do not freeze receipt of may require (Varicella- a skude ("touch prep") sample Chain of Custody zoster virus), and herpes Vesicles or pustules: Cell culture Refrigerate at 4-10 days See above re COC simplex virus. use scapel to open, 2-8C PCR performed at remove top of the CDC DO NOT ORDER vesicle or pustule THESE TESTS and place the skin IF PATIENT of the vesicle top FALLS INTO into a 1.5-2.0 ml HIGH RISK screw capped plastic CATEGORY. vial. REFER TO THE SMALLPOX Scab lesions: place Cell culture Refrigerate at 4-10 days See above re: COC HIGH RISK two scabs in each 2-8C PCR performed at CHART!! of two 1.5-2.0 ml CDC screw capped plastic vial. Blood: 10 cc of IFA test for detection Refrigerate at 2-24 hrs after See above blood collected in of specific antibodies 2-8C initial setup a serum separator in serum tube Environmental No method available at --- --- --- --- DPHL or CDC Food No method available at --- --- --- --- DPHL or CDC ****CDC Smallpox Draft Guide C, Part 1, Infection Control Measures for Healthcare and Community Settings, Guide F, Environmental Control f Smallpox Virus, http://www.bt.cdc.gov/agents/smallpox/infection-control/ ______________________________________________________________________________________________________________________ DISEASE/AGENT - VHF - Viral hemorrhagic fevers SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical Blood: 10 -12 cc Sent to CDC Decontaminate CDC will CDC requires of serum in a outside of tube, notify DPHL BioSafety Level 4 red/black top tube CDC will determine place in a double of results. facilities to be which tests to be leak proof container DPHL will used to test for performed then notify these viruses. submitter as DPHL does not have Proper PPE including soon as a BioSafety Level 4 gloves should be worn possible. Lab to test for while collecting specimen these viruses. All submitted specimens will therefore be referred to CDC for testing. Clinical specimens may require Chain of Custody. Environmental No method available at --- --- --- --- DPHL or CDC Food No method available at --- --- --- --- DPHL or CDC ___________________________________________________________________________________________________________________________ DISEASE/AGENT - Plague (Yersinia pestis) SPECIMEN SELECTION FROM SAMPLES TEST METHOD TRANSPORT/MEDIUM PROCESSING COMMENTS INSTRUCTIONS TIME Clinical **Bronchial/tracheal Culture: rule-out and Grown isolate on 24-72 hrs *Cultures from wash**: collect per culture confirmation agar plate sent at clinical specimens institution’s available on grown room temp should be performed procedures in an area organism. by the sentinel dedicated to collection laboratory if of respiratory possible specimens under isolation/containment PCR: available on Refrigerate at 8-24 hrs May require Chain circumstances, i.e., grown organism only Refrigerate at of Custody isolation chamber/ PCR requires ”bubble” culture confirmation *Sputum/throat:collect Culture: rule-out and Grown isolate on 24-72 hrs Minimal recovery-- routine throat culture culture confirmation agar plate sent at not recommended using a swab or collect available on grown room temp expectorated sputum organism. into a sterile, leak proof container. PCR: available on Refrigerate at 8-24 hrs May require Chain grown organism 2-8C of Custody PCR requires culture confirmation *Blood Culture:collect Culture: rule-out Grown isolate on 24-72 hrs Blood culture per institution’s and culture agar plate sent at is rarely positive procedure for routine confirmation room temp Blood culture blood culture. Blood available on cannot be culture is incubated grown organism. performed at DPHL by facility drawing the culture and PCR: available on Grown isolate on 4-8 hrs PCR requires screened for growth of grown organism agar plate sent at culture the organism. room temp confirmation *Nasopharyngeal swabs: Culture: rule-out Grown isolate on 24-72 hrs for PCR the swabs must and culture agar plate sent at be collected in a confirmation room temp sterile tube without available on transport media grown organism. PCR: available on <2hrs - room temp 8-24 hrs May require grown organism >2hrs - refrigerate Chain of Custody at 2-8°C PCR requires culture confirmation Environmental Powder, letter, etc.: Culture: available Use Dacron or 8-24 hrs Chain of Custody collect at least two polyester swabs or required non-cotton swabs, a culturette place in separate sterile containers. PCR: available on Use Dacron or 4-8 hrs for Chain of Custody original specimen polyester swabs, no organism required on and grown organisms culturettes 8-24 hrs for environmental environmental samples PCR requires culture confirmation Food Notification of DPHL Culture and PCR: Refrigerated samples 2-7 days Chain of Custody by DPH Epidemiology available & those collected at required Section is necessary Send all suspect room temp should be as soon as possible food in a leak frozen at 4C. Frozen PCR requires due to the large proof container. samples should be culture amount of media Minimum needed to transported on dry confirmation necessary to test: 10 grams ice. process food. Submitter must contact DPH Epidemiology Section concerning any suspected food related infections. *Cultures from clinical specimens should be performed by the sentinel laboratory if possible Revised 03/01/07