Procedures

WWARN is continually updating a library of standardised procedures that offer guidance in the execution of various activities in the fields of clinical, in vitro, pharmacology, data collection and molecular analysis for antimalarial drug resistance researchers. 

For resources that relate to other infectious and emerging diseases, visit iddo.org.

INV11
Piperaquine Survival Assays (PSA) to evaluate the in-vitro and ex-vivo susceptibility of Plasmodium falciparum to piperaquine
This procedure describes two assays to evaluate the susceptibility of Plasmodium falciparum to piperaquine.
Version: V1.0 Author: Benoit Witkowski & Didier Ménard, Institut Pasteur du Cambodge
CLI20; PHA11
Malaria Case Record Form (CRF)
Malaria Clinical Trial Case Record Form
Version: v1.0 Author: WWARN
CLI19
Obare Method Calculator
A tool to calculate the mean parasite density and assessing whether two blood samples are concordant
Version: v1.0 Author: Obare
EXT11
Guidelines for the Cryopreservation of field isolates
The purpose of this document is to standardize the methodology for cryopreservation of malaria parasites using glycerolyte
Version: 1.0 Author: Dr. Kesinee Chotivanich
INV10
Ring-stage Survival Assays (RSA) to Evaluate the In-Vitro and Ex-Vivo Susceptibility of Plasmodium Falciparum to Artemisinins
Evaluting Plasmodium falciparum susceptibility to artemisinins in laboratory strains and fresh isolates
Version: v1.2 Author: Institut Pasteur du Cambodge & National Institute of Allergy and Infectious Diseases/NIH
EXT10
Microscopy for the detection, identification and quantification of malaria parasites on stained thick and thin blood films in research settings
This manual was developed to guide a move towards common standards for undertaking and reporting research microscopy for malaria parasite detection, identification and quantification.
Version: 1.0 Author: Research Malaria Microscopy Standards Working Group
PHA10
Guidelines for the collection of samples for artemisinin pharmacology analysis
These guidelines aim to standardize the sample collection procedure during clinical studies of artemisinin antimalarial drugs
Version: 1.0 Author: Chris Lourens
PHA09
Guidelines for the collection of samples for primaquine / carboxyprimaquine pharmacology analysis
These guidelines aim to standardize the sample collection procedure during clinical studies of primaquine / carboxyprimaquine antimalarial drugs
Version: 1.0 Author: Chris Lourens
PHA08
Guidelines for the collection of samples for piperaquine pharmacology analysis
These guidelines aim to standardize the sample collection procedure during clinical studies of piperaquine antimalarial drugs
Version: 1.0 Author: Chris Lourens
PHA07
Guidelines for the collection of samples for mefloquine / carboxymefloquine pharmacology analysis
These guidelines aim to standardize the sample collection procedure during clinical studies of mefloquine / carboxymefloquine antimalarial drugs
Version: 1.0 Author: Chris Lourens

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48 results

Serious Adverse Event Management

Identifying, assessing and reporting serious adverse events during antimalarial drug efficacy trials
Version: v1.0 Author: WWARN CLI04 Participants: Clinical Scientific Group

Blood collection: peripheral venipuncture

Collecting peripheral blood samples
Version: v1.0 Author: WWARN CL108 Participants: Clinical Scientific Group

Weight Measurement

Measuring and recording patient weight in antimalarial drug efficacy trials
Version: v1.0 Author: WWARN CLI02 Participants: Clinical Scientific Group

Temperature Measurement: Digital Thermometer

Measuring and recording patient axillary body temperature in clinical trials
Version: v1.0 Author: WWARN CLI17 Participants: Clinical Scientific Group

Sample Preservation

Collecting and preserving DNA, RNA and parasite isolates from infected blood samples
Version: v1.5 Author: WWARN MOL01 Participants: Molecular Scientific Group

Collection of blood by finger prick and heel stick

Obtaining fingerprick and heel stick blood samples for preparing thick/thin blood smears and filter paper samples
Version: v1.2 Author: WWARN CLI07 Participants: Clinical Scientific Group

Temperature Measurement: Braun Thermoscan Thermometer

Measuring and recording patient tympanic membrane body temperature in clinical trials
Version: v1.2 Author: WWARN CLI16 Participants: Clinical Scientific Group

Respiratory Rate Measurement

Measuring and recording patient respiratory rate in antimalarial drug efficacy trials
Version: v1.0 Author: WWARN CLI01 Participants: Clinical Scientific Group

Leukocyte Depletion of Whole Blood Using CF11 Columns

Version: v1.3 Author: WWARN MOL02 Participants: Molecular Scientific Group

Collection of blood on filterpaper

Preparing and preserving dried blood spot samples for genetic analysis
Version: v1.2 Author: WWARN MOL03, CLI06 Participants: Molecular Scientific Group, Clinical Scientific Group

Database Construction and Security

Construction, security and backup of a clinical database
Version: v1.0 Author: WWARN PHA01 Participants: Pharmacology Scientific Group

Data Receipt, Entry and Validation

Receiving, tracking and validating data within a clinical trial
Version: v1.0 Author: WWARN PHA02 Participants: Pharmacology Scientific Group

Preparation of Culture Medium

Preparing complete medium for Plasmodium falciparum cultures
Version: v1.1 Author: WWARN INV02 Participants: In Vitro Scientific Group

Synchronisation of Plasmodium falciparum

Synchronising culture adapted Plasmodium falciparum isolates in ring stage
Version: v1.1 Author: WWARN INV04 Participants: In Vitro Scientific Group

Culture of Plasmodium falciparum blood stages

Methods for short- and long term culture of Plasmodium falciparum laboratory strains and fresh isolates
Version: v1.0 Author: WWARN INV01 Participants: In Vitro Scientific Group

DNA Extraction: Preparing Rapid Diagnostic Tests

Storing and processing malaria Rapid Diagnostic Tests (RDTs) for DNA extraction
Version: v1.1 Author: WWARN MOL06 Participants: Molecular Scientific Group

Quantification of Human and P. Falciparum DNA

Estimating Plasmodium falciparum and human DNA concentration in infected blood samples by fluorometer and real-time PCR
Version: v1.0 Author: WWARN MOL04 Participants: Molecular Scientific Group

Copy number estimation of the Plasmodium falciparum pfmdr1 gene

Estimating pfmdr1 gene copy number by real-time PCR
Version: v1.1 Author: WWARN MOL05 Participants: Molecular Scientific Group

Preparation of drug plates for susceptibility assays

Preparation of drug dilutions in dry or fresh plates for drug susceptibility assays
Version: v1.2 Author: WWARN INV03 Participants: In Vitro Scientific Group

Estimation of Plasmodium falciparum drug susceptibility by the 3H-hypoxanthine uptake inhibition assay

Drug susceptibility assay measuring radioactive hypoxanthine uptake in laboratory strains and fresh isolates.
Version: v1.0 Author: WWARN INV07 Participants: In Vitro Scientific Group

Blood Smear Preparation

Preparing thick and thin blood smears with Giemsa staining
Version: v1.0 Author: WWARN CLI10 Participants: Clinical Scientific Group

Adverse Event Management

Identifying, assessing and reporting adverse events during antimalarial drug efficacy trials
Version: v1.1 Author: WWARN CLI05 Participants: Clinical Scientific Group

Estimation of Plasmodium falciparum drug susceptibility ex vivo by HRP2 ELISA

Drug susceptibility assay measuring histidine-rich protein production by ELISA in fresh isolates
Version: v1.0 Author: WWARN INV09 Participants: In Vitro Scientific Group

Estimation of Plasmodium falciparum drug susceptibility by the SYBR® Green assay

Drug susceptibility assay measuring SYBR®Green I fluorescence in laboratory strains and field isolates
Version: 1.0 Author: WWARN INV08 Participants: In Vitro Scientific Group

Collection of blood on filterpaper

Preparing dried blood spots by finger prick
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT01 Participants: Molecular Scientific Group, Clinical Scientific Group

DNA extraction by Chelex

Extracting Plasmodium spp. DNA from dried blood spots by Chelex 100
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT02 Participants: Molecular Scientific Group

DNA Extraction by QIAamp DNA Mini Kit

Extracting Plasmodium spp. DNA from dried blood spots by QIAamp DNA Mini Kit
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT03 Participants: Molecular Scientific Group

msp1 PCR

PCR amplification of the polymorphic marker msp1 to distinguish recrudescences from reinfections
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT05 Participants:

msp2 PCR

PCR amplification of the polymorphic marker msp2 to distinguish recrudescences from reinfections
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT09 Participants: Molecular Scientific Group, Clinical Scientific Group

glurp PCR

PCR amplification of the polymorphic marker glurp to distinguish recrudescences from reinfections
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT06 Participants: Molecular Scientific Group, Clinical Scientific Group

Manual size estimation of msp1, msp2 and glurp PCR products on gel

Manually determining the size of Plasmodium falciparum polymorphic markers to distinguish recrudesent from reinfecting infections in clinical trials
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT08 Participants: Clinical Scientific Group, Molecular Scientific Group

Agarose Gel Electrophoresis of msp1, msp2 and glurp

Separating and visualising amplified PCR products of msp1, msp2 and glurp
Version: 1.0 Author: Mekong Molecular Surveillance Network EXT07 Participants:

Ring-stage Survival Assays (RSA) to Evaluate the In-Vitro and Ex-Vivo Susceptibility of Plasmodium Falciparum to Artemisinins

Evaluting Plasmodium falciparum susceptibility to artemisinins in laboratory strains and fresh isolates
Version: v1.2 Author: Institut Pasteur du Cambodge & National Institute of Allergy and Infectious Diseases/NIH INV10 Participants: In Vitro Scientific Group

PCR and sequencing for genotyping of candidate Plasmodium falciparum artemisinin resistance SNPs in the Kelch 13 gene

Genotyping of SNPs in the Kelch 13 gene (PF3D7_1343700), associated with artemisinin resistance
Version: 1.0 Author: D. Menard & F. Ariey Institut Pasteur MOL08 Participants: Molecular Scientific Group

Data recording and internal quality checks

Processes and instructions for recording and validating data (source and case record form) generated during a clinical trial
Version: v 1.0 Author: WWARN PHA03 Participants: Pharmacology Scientific Group

AE_SAE Improvement

Version: 1.0 Author: WWARN CLI18 Participants: Clinical Scientific Group

Microscopy for the detection, identification and quantification of malaria parasites on stained thick and thin blood films in research settings

This manual was developed to guide a move towards common standards for undertaking and reporting research microscopy for malaria parasite detection, identification and quantification.
Version: 1.0 Author: Research Malaria Microscopy Standards Working Group EXT10 Participants:

Guidelines for the collection of samples for pyronaridine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of pyronaridine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA04 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for amodiaquine / desethylamodiaquine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of pyronaridine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA05 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for lumefantrine / desbutyl-lumefantrine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of lumefantrine / desbutyl-lumefantrine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA06 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for mefloquine / carboxymefloquine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of mefloquine / carboxymefloquine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA07 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for piperaquine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of piperaquine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA08 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for primaquine / carboxyprimaquine pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of primaquine / carboxyprimaquine antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA09 Participants: Pharmacology Scientific Group

Guidelines for the collection of samples for artemisinin pharmacology analysis

These guidelines aim to standardize the sample collection procedure during clinical studies of artemisinin antimalarial drugs
Version: 1.0 Author: Chris Lourens PHA10 Participants: Pharmacology Scientific Group

Guidelines for the Cryopreservation of field isolates

The purpose of this document is to standardize the methodology for cryopreservation of malaria parasites using glycerolyte
Version: 1.0 Author: Dr. Kesinee Chotivanich EXT11 Participants:

Obare Method Calculator

A tool to calculate the mean parasite density and assessing whether two blood samples are concordant
Version: v1.0 Author: Obare CLI19 Participants:

Malaria Case Record Form (CRF)

Malaria Clinical Trial Case Record Form
Version: v1.0 Author: WWARN CLI20; PHA11 Participants: Pharmacology Scientific Group

Piperaquine Survival Assays (PSA) to evaluate the in-vitro and ex-vivo susceptibility of Plasmodium falciparum to piperaquine

This procedure describes two assays to evaluate the susceptibility of Plasmodium falciparum to piperaquine.
Version: V1.0 Author: Benoit Witkowski & Didier Ménard, Institut Pasteur du Cambodge INV11 Participants: