TestpolEng 2

Public Health Service
DEPARTMENT OF HEALTH & HUMAN SERVICES
CDC Dengue Branch

Division of Vector-Borne Diseases
National Center for Emerging and
Zoonotic Infectious Diseases
1324 Caada Street
San Juan, Puerto Rico 00920-3860
Tel: 787-706-2399; Fax: 787-706-2496
Criteria for the processing of dengue samples at the

CDC Dengue Branch, San Juan, Puerto Rico
The CDC Dengue Branch provides dengue testing free of cost to submitting physicians, state and private
laboratories. To adequately determine what laboratory tests are necessary, correctly provide a clinical
diagnosis and to assure that test results are received by the requesting physician, the following
information is essential:
Complete name, age and sex of the patient

Home address
Date of onset of symptoms
Date that sample was obtained
If the patient traveled prior to disease presentation, travel destination and dates need to be
included.
Complete name and mailing address of the physician, laboratory, clinic or hospital that the
result should be sent to.
Samples without the above-mentioned information, written with illegible handwriting or with more
than a month from date of sample collection to date of arrival at CDC, will not be tested.
In circumstances of a dengue outbreak or cases of severe disease such as hemorrhagic manifestations,
the CDC Dengue Branch will promptly analyze samples received with the minimum above-mentioned
information. If number of specimens exceeds laboratory capacity, testing may be prioritized in the
following order:
cases in intensive care,

hospitalized cases (with thrombocytopenia, hemorrhage, shock or hemoconcentration
fatal cases ),
all other DF cases.
Case Notification and Shipment of Samples of Suspected Dengue Cases

Dengue is an acute febrile illness. In addition to fever, headache, pain behind the eyes, body aches, rash,
nausea and vomiting are common symptoms of dengue. Other variable symptoms can include
hemorrhagic manifestations (such as a petechial rash) or hemoconcentration, shock or coma. Serum
samples from dengue suspected cases should be obtained as early as possible and submitted to State
Health Department laboratories. Samples obtained during the first 5 days of illness (acute samples) are
likely to contain virus; whereas samples collected after 5 days of illness (convalescent samples) may
contain antibodies against the virus. Samples can be submitted to the CDC Dengue Branch; following
reporting to the State Laboratory.
Instructions for obtaining and handling samples:
1. Once there is a clinical diagnosis of suspected dengue, take a blood sample (see #3 to 5) and fill out
the Dengue Case Investigation Form (see copy attached). With this Form you comply with the legal
reporting requirement. These forms can be obtained from the CDC Dengue Branch website
http://www.cdc.gov/dengue/resources/dengueCaseReports/DCIF_English.pdf or the Puerto Rico
Department of Health website
http://www.salud.gov.pr/Datos/VDengue/Documents/InformeInvestigacionCD.pdf. Forms can also
be photocopied without restriction.
2. It is important to fill out the Dengue Case Investigation Form in a clear and complete manner. The
information received on each case (especially the date of onset of symptoms and date of sample
collection) is crucial to determine the appropriate diagnostic test and interpret the laboratory
results. Furthermore, a complete address makes it possible to identify regions of transmission
where control measures should be implemented. Samples without the above-mentioned
information, or written in illegible handwriting or with more than a month from date of collection
to date of arrival at CDC, will not be tested.
3. Serum samples are preferred for most dengue tests. The blood sample should be taken in a red-top
or tiger-top tube. Citrate (collected in yellow top tubes) and heparin plasma (green top tubes) can
be tested by RT-PCR. Violet-top (with EDTA) is not recommended for RT-PCR testing. Violet and or
green-top tubes should not be used for serology testing (convalescent sample). Please refer to
collection devices manufacturer instructions for more details. After blood is allowed to clot,
separate serum by centrifugation and keep serum refrigerated at 4 oC or frozen at -20 oC (preferred).
4. Specimen should be sent to the CDC-Dengue Branch (address mentioned on the Dengue Case
Investigation Form).
For samples from the USA or abroad, we recommend to freeze the serum immediately after
separated and to send on dry ice. If dry ice is not available, we recommend that the serum is kept
refrigerated and delivered to the CDC Dengue Branch in cold packs. The case investigation forms
and the acute serum sample should reach CDC Dengue Branch as soon as possible following
collection; there is no need to wait until the convalescent sample is obtained. Most dengue
diagnostic tests practiced at the CDC-Dengue Branch have been validated in serum and plasma
samples. Therefore, plasma separated in lavender (EDTA) or green (heparin) top tubes are
acceptable for PCR testing only, not for serology testing. These samples should also be stored and
sent to the CDC Dengue Branch in the conditions specified for sera.
5. To diagnose dengue, the laboratory requires a serum sample obtained during the acute phase of the
infection (DPO=0-5). If this sample is negative, then a second convalescent serum sample (that can
be taken from day 6 after the onset of symptoms) is required to confirm the case. Informing the
patient about the importance of returning for a second sample, and providing an appointment for a
specific day and hour, will increase the probability of obtaining the second sample.
6. Acute-phase samples (taken on or before day 5 after onset of symptoms), will be tested by RT-PCR
for virus detection and convalescent-phase samples (taken on or after day 6 after beginning of
symptoms) will be tested for anti-dengue IgM antibodies by enzyme-linked immunosorbent assays
(ELISA). Differential diagnosis for dengue and WN virus is available; but these tests need to be
requested according to clinical presentation.
Type of
sample
Acute
Convalescent
Interval since
the onset of symptoms
Type of analysis
until day 5
RT-PCR
6 or more days
Serology
Samples taken on days 4 and 5 of illness are of low yield for isolation as well as serology.
WHENEVER THERE IS A HOSPITALIZED SEVERE CASE, PLEASE INDICATE on THE CASE INVESTIGATION
FORM.
7. Reports will be sent to the physician (if the return address has been indicated) with the results of
positive or negative cases. In cases with negative RT-PCR results, we will wait for a convalescentphase sample before reporting a result. If a convalescent sample is not obtained, the case will be
considered indeterminate.
8. Results will be reported only to the laboratory or the physician who sent the sample (or authorized
personnel).
Mailing address:
Dengue Branch, CDC
1324 Caada St.
Puerto Nuevo
San Juan, Puerto Rico 00920-3860
Points of contact:
Dr. Jorge Muoz ckq2@cdc.gov for molecular testing
Dr. Elizabeth Hunsperger enh4@cdc.gov for serology testing
Tel. (787) 706-2399

Fax (787) 706-2496
CHECK LIST FOR OBTAINING AND SHIPPING DENGUE DIAGNOSTIC SAMPLES
[ ] Sample
Type of sample
Interval since date
Type of analysis
Acute
of onset of symptoms
Up to 5 days
RT-PCR
Convalescent
6 or more days
Serology
[ ] Form - "Dengue Case Investigation Form"

Can be obtained from the CDC Dengue Branch in San Juan or Internet:
http://www.cdc.gov/dengue/resources/dengueCaseReports/DCIF_English.pdf Please indicate on
the sheet if the case is hospitalized. If it is a very severe case, indicate so on the "Comments"
section.
Only the samples received with the information requested below, and written in a legible
manner, will be analyzed:
Complete name, age, and sex of patient
Home address
Date of onset of symptoms
Date sample was obtained
Complete name and mailing address of the physician, laboratory, clinic, or hospital
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Tube Red, tiger-top or green top (not violet).

Labeling - Tube and case form must agree (indicate the same name of the case).
Volume - 2 cc. (ml.) of centrifuged serum or plasma
Storage - On ice or in a refrigerator until it is delivered to the CDC Dengue Branch.
Time of shipment - Not to exceed a month after taking the sample
Way of shipment - Check with your local Department of Health.
Reasons for REJECTING samples:
Samples without form, form without sample
Incomplete or illegible form especially regarding date of onset of symptoms, date of

sample collection
Hemolyzed or contaminated samples are not acceptable for serology testing
Samples received more than a month after onset of illness

Dengue Branch, CDC
1324 Calle Caada
Puerto Nuevo
San Juan, PR 00920-3860
January 19, 2012

Tel. (787) 706-2399
Fax (787) 706-2496

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Public Health Service

DEPARTMENT OF HEALTH & HUMAN SERVICES

CDC Dengue Branch

Criteria for the processing of dengue samples at the

Complete name, age and sex of the patient

cases in intensive care,

Case Notification and Shipment of Samples of Suspected Dengue Cases

Tel. (787) 706-2399

CHECK LIST FOR OBTAINING AND SHIPPING DENGUE DIAGNOSTIC SAMPLES

Interval since date

[ ] Form - "Dengue Case Investigation Form"

Tube Red, tiger-top or green top (not violet).

Reasons for REJECTING samples:

Samples without form, form without sample

Incomplete or illegible form especially regarding date of onset of symptoms, date of

Hemolyzed or contaminated samples are not acceptable for serology testing

Samples received more than a month after onset of illness

January 19, 2012

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