Download impaact 2010 (vested)

IMPAACT 2010 (VESTED)
Laboratory Considerations
Laboratory Technologist: Sikhulile Moyo
IMPAACT Lab Center Rep: William Murtaugh
QUICK REVIEW OF STUDY ACTIVATION:
LABORATORY REQUIREMENTS
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IMPAACT Laboratory Center (LC) approval of local
laboratory site readiness, based on confirmation of the
following:
All laboratories
• Confirmation of maternal and infant HIV testing
algorithms
• Confirmation of access to Abbott Real-Time HIV-1
RNA PCR assay at local VQA-certified laboratory
• Confirmation of current IATA training certificates for
at least two individuals
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IMPAACT Laboratory Center (LC) approval of local
laboratory site readiness, based on confirmation of the
following:
Additionally for non-US site laboratories
• DCLOT approval of laboratory readiness based on:
– Completion of relevant IMPAACT Critical Items from DAIDS Audit
Action Plan, if applicable
– Approval of protocol analyte list (PAL)
– Confirmation of acceptable EQA status (pSMILE, VQA, IQA) for
protocol analytes for both primary and backup laboratories
– Submission of current maternal and pediatric reference ranges
(must be reviewed, approved, and signed by Laboratory Medical
Director)
– Have made significant progress toward obtaining required
permits and M/STAs for shipping specimens to the University of
Washington
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Accessing Laboratory Related
Documents
Protocol Specific Webpage:
• Laboratory Processing Chart (LPC)
• Manual of Procedures (MOP)
Email Communication with ILC Rep:
• Protocol Analyte List (PAL)
IMPAACT Laboratory Center webpage:
http://impaactnetwork.org/aboutus/LabCenter/LabGuidance.htm
• HIV NAT Assay requirements
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REVIEW OF THE
LABORATORY PROCESSING CHART (LPC)
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LPC includes information on…
•
•
•
•
•
•
Blood volumes
tube types
LDMS test codes
Relevant Case Report Forms (eCRF)
Collection, processing, storage requirements
Shipping details
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LPC Sections
• Protocol Required Non-Standard Reagents and Supplies
• Section A (1a-4a): Maternal Laboratory Processing Instructions
–
–
–
–
1a: Antepartum, Delivery and Postpartum SoE
2a: Safety/Clinical Laboratory Evaluations
3a: Specimen Processing
4a: Evaluations by Visit
• Section B (1b-4b): Infant Laboratory Processing Instructions
–
–
–
–
1a: Delivery and Postpartum SoE
2a: Safety/Clinical Laboratory Evaluations
3a: Specimen Processing
4a: Evaluations by Visit
• Section 5: Shipping Instructions
• Section 6: Revision History
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Protocol Required Non-standard
Reagents and Supplies
• There are no unusual reagents and supplies required
for this study.
• However, sites should always ensure that they have
all the required tube volumes and tube types by the
time of study activation.
– It is essential to use the correct tube volume and tube type
so as not to compromise sample integrity and laboratory
results.
• If you have difficulty obtaining tube supplies, contact
the IMPAACT Laboratory Center Representative
(William Murtaugh)
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MATERNAL LABORATORY EVALUATIONS
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Prioritization of Blood Collection and
Processing for Short Draws: MATERNAL
IMPAACT 2010 complies with NIH recommendations to limit adult blood collection to 10.5 mL/kg in a single day
and 550 mL in any 8-week period. In the event that maternal blood collection must be limited, refer to the blood
draw priority list below.
Visit
Order of blood
collection
Purpose
2
SST or NON Confirmatory HIV testing (if needed at
or EDTA
Screening Visit)
EDTA
HIV-1 RNA
3
SST or NON Chemistries (ALT, AST, creatinine)
4
SST or NON
Hepatitis B surface antigen (required at Entry
Visit only)
5
EDTA
Hematology (complete blood count)
6
EDTA
CD4+ cell count
7
EDTA
Storage (plasma, cell pellets, and serum)
1
Screening
through Week 50
Tube type
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Section 2a: Safety/Clinical Laboratory Evaluations – MATERNAL VISITS
Defer to local clinical specimen collection guidelines for tube types and collection volumes as needed.
Evaluation
Tube Type
Confirmatory HIV testing
SST or NON or
EDTA
Hepatitis B Surface Antigen
(HBsAg) testing
Hematology
SST or NON
EDTA
Chemistries
SST or NON
CD4+ cell count
EDTA OR DPE
Urine Pregnancy Testing
Urine
Serum Pregnancy Testing
SST or NON
HIV-1 RNA PCR
(Abbott RealTime Viral Load
Assay)
EDTA
Tests
eCRF(s)
Complete HIV testing if needed to meet criteria specified in
protocol Section 4.1.2
N/A
Laboratory
Test Results
(Chem/Hem)
Hepatitis B surface antigen
Complete blood count (CBC; to include red blood cell count,
hemoglobin, hematocrit, white blood cell count, white blood
cell differential, absolute neutrophil count, and platelet
count)
ALT, AST, creatinine
Note: Creatinine Clearance (CrCl) may be calculated by clinic
or laboratory staff, and should be calculated based on
creatinine results using the Cockcroft-Gault formula
CD4+ absolute cell counts
 HCG (pregnancy test)
Test used must have sensitivity of 25mIU or less.
 HCG (pregnancy test)
Test used must have sensitivity of 25mIU or less.
The total blood volume listed in the SoE at the Screening and
Week 50 Visits accommodates collection of 1mL of blood if
needed for serum pregnancy testing.
HIV-1 RNA using the Abbott platform in a VQA-certified lab
(all sites).
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Laboratory
Test Results
(Chem/Hem)
Laboratory
Test Results
(Chem/Hem)
Lymphocyte
Subsets
Pregnancy
Test
Pregnancy
Test
Specimen
Tracking
HIV-1 RNA
Plasma Viral
Load
Safety/Clinical Lab Considerations
Confirmation of HIV Infection (if required at screening):
• Testing algorithm must adhere to protocol section 4.1.2 and be approved
by the IMPAACT Lab Center
Chemistries:
• Creatinine Clearance (CrCl) may be calculated by clinic or laboratory staff,
and should be calculated based on creatinine results using the CockcroftGault formula
Serum pregnancy testing (if required at screening):
• The total blood volume listed in the SoE at the Screening and Week 50
Visits accommodates collection of 1mL of blood if needed for serum
pregnancy testing.
Maternal HIV RNA Testing:
• Real-time testing is required to be performed on the Abbott platform
(Real-Time HIV-1 viral load assay) by a VQA-approved laboratory
• Will be confirmed on Protocol Analyte List (PAL)
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Confirmation of Maternal Infection
Sample #1
Sample #2
• Two different rapid antibody
tests
• Rapid antibody test for a total
of three different rapid tests
• One EIA or WB or
immunofluorescence or
chemiluminescence test
• One EIA or WB or
immunofluorescence or
chemiluminescence test
• One HIV DNA PCR
• One HIV DNA PCR
• One quantitative HIV RNA PCR • One quantitative HIV RNA PCR
(result above LOD)
(result above LOD)
• One qualitative HIV RNA PCR
• One qualitative HIV RNA PCR
• One total nucleic acid test
• One total nucleic acid test
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Confirmation of Maternal Infection
• If both samples are tested using antibody tests, at least one
must be tested in a laboratory setting that operates according
to GCLP guidelines and participates in an appropriate EQA
program
• If nucleic acid testing is used, at least one test must be
performed in a CLIA-certified (US sites) or VQA-certified
(non-US sites) laboratory
• For tests performed in other settings, adequate source
documentation including the date of specimen collection, date
of testing, test performed, and test result must be available
• FDA-approved methods should be used when possible, and
when a combination of three rapid tests are performed, at
least one must be FDA approved
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MATERNAL SPECIMEN PROCESSING
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Section 3a: Specimen Processing – MATERNAL VISITS
Refer to Section 4 for collection volumes.
Evaluation
Plasma For HIV-1 RNA
(Abbott RealTime
Viral Load Assay)
Perform in real time, store
residual plasma
Plasma storage for
HIV-1 RNA at centralized lab
Plasma storage for
ARV drug resistance testing
Tube Type
EDTA
EDTA
EDTA
Special
eCRF(s)
Collection Notes
Invert 8-10
times gently.
Invert 8-10
times gently.
Invert 8-10
times gently.
Processing
Shipping
Spin blood at 400 x g for
Send to VQA certified lab.
10 min. Remove plasma
Specimen
Store residual plasma at
and respin at 800 x g for
Tracking
site until requested by
10 min. Freeze residual
protocol team.
plasma at -70°C or
colder.
Spin blood at 400 x g for
Specimen 10 min. Remove plasma
Tracking and respin at 800 x g for
10 min. Freeze plasma at
-70°C or colder.
Spin blood at 400 x g for
Specimen 10 min. Remove plasma
Tracking and respin at 800 x g for
10 min. Freeze plasma at
-70°C or colder.
Store at site until
requested by
protocol team for batch
testing at centralized
virology lab.
Refer to Section 5 for
shipping details.
Store at site until
requested by
protocol team for
batch testing at
centralized virology
laboratory.
Refer to Section 5 for
shipping details.
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Screening Visit (window: up to -14d)
LDMS VISIT CODE = 0/SCR
Evaluation
Specimen
eCRF(s)
Aliquots
LDMS Code
Special Notes
Confirmatory
HIV testing
SST or NON or
EDTA
0-6 mL
N/A
N/A
N/A
Complete HIV testing as needed per protocol
Sections 4.1.2 and 6.1.
Hematology
(complete blood
count)
EDTA
3 mL
Laboratory
Test Results
(Chem/Hem)
N/A
N/A
Send to local lab .
Send to local lab.
Chemistries (AST,
ALT, creatinine, CrCl)
Stored plasma
for ARV drug
resistance testing
Confirmatory
serum or urine
pregnancy test
SST or NON
4 mL
Laboratory
Test Results
(Chem/Hem)
N/A
2 x 1.0 mL
EDTA
6 mL
EDTA
1 mL
Urine
5 mL
Specimen
Tracking
Pregnancy
Test
Store residual:
1 x 0.5 – 1.0 mL
N/A
N/A
BLD/EDT/PL2
(BLD/DPE/PL2)
Creatinine Clearance (CrCl) may be calculated by
clinic or laboratory staff, and should be calculated
based on creatinine results using the CockcroftGault formula
Specimens collected and stored at screening for
participants who do not subsequently enroll should
be destroyed.
For enrolled participants, store aliquots on site until
requested by protocol team. Refer to Section 5 for
shipping details.
N/A
If needed per protocol Sections 4.1.6 and 6.1.
Send to local lab.
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Maternal Specimen Processing
Plasma: Residual plasma should be stored wherever possible.
• Abbott RealTime HIV-1 viral load
– Local VQA-approved laboratory
• Plasma storage for centralized HIV-1 viral load testing
– Store until requested to batch ship to U. Washington
• Plasma storage for ARV Drug Resistance testing
– Store until requested to batch ship to U. Washington
– If participant is not enrolled, stored plasma sample collected at
screening should be destroyed
• Plasma Storage for markers of adverse pregnancy outcomes
– Store at site until requested by protocol team
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Maternal Specimen Processing
Cell Pellet (non-viable PBMCs):
• Storage for markers of adverse pregnancy outcomes
• Site should follow the Cross-Network PBMC Processing SOP
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Maternal Specimen Processing
Serum:
• Zika diagnostic testing (if indicated)
• Store at site until requested by protocol team
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Maternal Specimen Processing
Hair Storage:
• PK testing
• Collection instructions can be found in the Manual of
Procedures (MOP)
• Hair sample should be stored at room temperature
without exposure to light
• Ship at request of protocol team to University of
California, San Francisco
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Maternal Specimen Processing
Breast Milk:
• Store at site until requested for batch testing at
centralized pharmacology lab (University of
California, San Diego)
– Whole Milk
– Skim Milk
– Dry cell pellet
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Breast Milk Collection and Processing
• Collected in 50mL conical centrifuge tube
• Keep breast milk as cold as possible through ALL
processing steps. Send to local processing lab on wet
ice or cold packs.
• Complete processing within 6 hours of collection. If
processing occurs outside this window, note this in
the LDMS.
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Breast Milk Collection and Processing
Whole Milk:
• Vortex 50 mL conical centrifuge tube
• Making at least 6x1mL aliquots in 2 mL cryovials.
• Freeze at -70°C or colder.
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Breast Milk Collection and Processing
Skim Milk:
• Transfer remaining whole milk to a 15mL conical
centrifuge tube.
• Centrifuge sample at 400xg for 20 min. Use sterile
spatula or wide tip pipette to remove lipid layer.
• Aliquot 4-6 x 1mL of skim milk, leaving approximately
0.5mL skim milk remaining. Do not disturb the pellet.
• Freeze skim milk aliquots at -70°C or colder.
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Breast Milk Collection and Processing
Cell Pellet:
• Add 1.0mL PBS to remaining skim milk
• Resuspend the pellet.
• Transfer to to 2mL centrifuge tube and spin at
2000xg for 3 minutes.
• Remove liquid without disturbing the pellet.
• Store dry cell pellet at -70°C or colder.
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Breast Milk Processing
Lab Processing
Clinic
20mL
Breast Milk
Tube Type
BMK
collected
in 50mL
conical
centrifuge
20mL
Breast Milk
Store at
4°C within
10 min of
collection
Send to
local lab
on wet ice
or cold
packs.
Whole
Milk:
Vortex
(5-10s)
6 x 1.0mL
cryovials
(Store at
-70C)
Centrifuge
remaining
Sample at
400xg for
20 mins.
Use Sterile
spatula or
wide tip
pipette to
remove
lipid layer
Skim
Milk:
4-6 x
1.0mL
cryovials
(Store at
-70C)
Approx.
0.5mL
skim milk
remaining
Add
1.0mL PBS
and
resuspend
the pellet
Transfer
to 2mL
centrifug
e tube
and spin
at
2000xg
for 3
minutes
Remove
liquid
without
disturbin
g the
pellet
and store
at -70C
or colder
Keep breast milk as cold as possible through ALL processing steps. Process within 6 hours of collection (>6 hrs. Note in LDMS)
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Maternal Specimen Processing
Urine storage:
• Markers of Renal Toxicity
• Keep specimen as cold as possible and complete
processing within 6 hours of collection. If processing
occurs outside this window, note this in the LDMS.
• Send to local processing lab on wet ice or cold packs.
• Aliquot and freeze at -70C or colder
• Store until requested by protocol team to ship to
designated testing lab
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INFANT
LABORATORY EVALUATIONS
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Prioritization of Collection of Blood Collection
and Processing for Short Draws: INFANT
IMPAACT 2010 complies with NIH recommendation to limit infant blood collection to
5 mL/kg in a single day and 9.5 mL/kg in any 8-week period. In the event that infant
blood collection must be limited, refer to the blood draw priority list below.
Order of Blood collection Tube type
Purpose
1
EDTA
HIV NAT
2
NON or SST
Chemistries (ALT, creatinine, ALT)
3
EDTA
Hematology (Complete Blood Count)
4
NON or SST
Serum storage
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INFANT SPECIMEN PROCESSING
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Infant Specimen Processing
HIV Nucleic Acid Testing:
• Performed locally for infants with unconfirmed infection
or who are uninfected
• Processing will be based on blood product required for
lab specific assay (two options):
– Whole blood (BLD)
– Whole blood pellet (PER)
– Plasma (PL2)
• All remaining aliquots and remnant blood products
should be frozen and stored until requested by the
protocol team
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Infant Specimen Processing
HIV Nucleic Acid Testing:
• At Infant Week 6 Visit, an additional 1mL of blood is
drawn for plasma PK testing (4mL total blood draw).
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Infant Specimen Processing
HIV Nucleic Acid Testing:
• For infants with confirmed HIV-1 infection:
– No additional diagnostic HIV tests will be performed
– Sample volume specified in the SoE will still be collected
– Sample will be processed, and stored for ARV Drug
Resistance testing
• Plasma storage
• Cell pellet storage
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Infant Specimen Processing
Serum:
• Zika diagnostic testing (if indicated)
• Store at site until requested by protocol team
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Infant Specimen Processing
Hair Storage:
• PK testing
• Collection instructions can be found in the Manual of
Procedures (MOP)
• Hair sample should be stored at room temperature
without exposure to light
• Ship at request of protocol team to University of
California, San Francisco
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ACTG/IMPAACT Laboratory Manual, Shipping Information and other useful information:
http://www.hanc.info/labs/labresources/Pages/informationActgImpaactLabs.aspx
NOTE: Please contact the laboratories prior to shipment to ensure they can receive the samples at the designated time. Do not ship until you receive
approval from the receiving lab. Ensure that all import permits are up to date and accurate prior to shipping samples.
VIROLOGY
Centralized HIV Viral Load testing
University of Washington
Retrovirus Lab
Attn: Joan Dragavon and Socorro Harb
HMC / R&T BLDG
300 Ninth Ave. RM 726
Seattle WA 98104
Email: [email protected]
Phone:206-897-5210
Fax: 206-897-5237
LDMS lab code: 15
PHARMACOLOGY
Plasma and Breast Milk for
ARV Drug Levels
IMPAACT Pharmacology
Specialty Laboratory at
University of California, San
Diego (UCSD)
Steven S. Rossi, Ph.D. or
Rowena Espina MT
Pediatric Pharmacology and
Antiviral Assay Laboratory
ATTN: Steve Rossi, Ph.D or
Rowena Espina MT
Centralized ARV drug resistance
212 Dickinson Street
testing
BLDG CTF-B, Room 112
Seattle Children’s Research Institute
San Diego, CA 92103-0808
Attn: Ingrid Beck or Sheila Styrchak
Phone: 619-543-5293
1900 9th Ave, Seattle WA 98101
Fax: 619-543-5422
email:
Email: [email protected]
frenkellabshipments@seattlechildrens Laboratory Contact: Rowena
.org
Espina
Phone:206-884-7201
Phone: 619-543-5293
Fax: 206-884-7311
Email: [email protected]
LDMS lab code: 238
LDMS lab code: 196
PHARMACOLOGY
Hair for ARV Drug Levels
Dr. Monica Gandhi
Attention: Karen Kuncze
c/o VESTED study (Shahin
Lockman)
Room S-907
513 Parnassus Ave
Medical Sciences Building, S864
San Francisco, CA 94143,USA
Email:
[email protected]
Email:
[email protected]
Phone: (415) 502-1446
Phone: 415 476 9960;
Fax number: 415-476-6770
LDMS lab code: 607
MARKERS OF
ADVERSE
PREGNANCY
OUTCOMES
PASS THROUGH SHIPPING
Biomedical Research Institute
9410 Key West Avenue
First Floor
Rockville, MD 20850
USA
Phone : (301) 881-7636
Fax: (301) 770-9811
Email:
[email protected]
LDMS lab code: 999
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Section 5: Shipping Instructions
• All shipping to occur upon request of the protocol team
• Laboratories should ship specimens directly to
specialized testing laboratories whenever possible.
• If your country or institution requires S/MTA, begin this
process as soon as possible to avoid shipping delays.
– It is the responsibility of the shipping and recipient laboratories
to complete paperwork and execute an agreement.
– Copies of signed MTAs must be made available to IMPAACT Lab
Center Representative
• For sites that are unable to ship directly, follow Cross
Network BRI pass through shipping instructions
39
Shipping Instructions (continued)
• Hair PK shipments must include certification
statement document from UCSF
• Document is available on the protocol specific
webpage under “Study Implementation Documents”
40
Section 6: Revision History
Version & Date
(dd/mmm/yy)
Comments
41
Questions?
Overall support for the International Maternal Pediatric Adolescent AIDS Clinical Trials (IMPAACT) Network was provided by the
National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH) under Award Numbers
UM1AI068632 (IMPAACT LOC), UM1AI068616 (IMPAACT SDMC) and UM1AI106716 (IMPAACT LC), with co-funding from the Eunice
Kennedy Shriver National Institute of Child Health and Human Development (NICHD) and the National Institute of Mental Health
(NIMH). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
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