Validation of RealAmp Method for the Diagnosis of Malaria in Endemic Areas of Brazil

• Sponsor: Centers for Disease Control and Prevention (U.S. Fed)
• Overall Status: Completed
• CT.gov ID: NCT02371395
• Collaborator: Evandro Chagas Institute of Clinical Research (Other)
• Enrollment: 1,000
• Location: 1
• Actual Duration (Months): 5
• Patients Per Site Per Month: 201.6

Study Details

Study Description

Brief Summary

Worldwide, approximately 2 billion people live in areas at risk for malaria with morbidity surpassing 250 million cases, with approximately 800,000 deaths, per year. Of the four species of malaria parasites that cause human infection, P. falciparum is responsible for the majority of severe malaria cases followed by P. vivax. Early and accurate diagnosis is essential for prompt and correct treatment, which can reduce the death rate and interrupt transmission. Currently, conventional methods for the diagnosis of malaria include microscopic examination of thin and thick blood smears and rapid diagnostic tests (RDTs). Light microscopy in practice typically detects parasitemia as low as 100 parasites/µl and it can differentiate species. The advantage of microscopy includes the ability to estimate parasitemia, the possibility to identify parasite stages, including gametocytes, and its low cost. However, this method is labor intensive, difficult to standardize, and requires well-trained microscopists. The majority of RDTs are based on detection of P. falciparum histidine-rich protein 2 (HRP-2) antigen and do not detect all malaria species. RDTs that detect lactose dehydrogenase (LDH) and aldolase generally broadly react with all four species of malaria parasites and therefore cannot differentiate among the species although efforts are underway to improve their performance for species detection. In settings where multiple malaria species co-circulate, molecular methods may be more reliable than microscopy and RDTs in accurately diagnosing the species of malaria parasites with low parasitemias. However, conventional molecular methods, such as nested polymerase chain reaction (nested PCR) or real-time PCR, are technically challenging and resource intensive and are generally restricted to reference laboratories due to the need for well-equipped laboratories. Recently, new molecular methods that can be used in field settings have been developed and this opens up new opportunities for exploring molecular tools for malaria diagnosis in endemic countries. With the objective of facilitating use of molecular tools for malaria control programs, the malaria laboratory at the Centers for Disease Control and Prevention (CDC) in Atlanta, USA developed a simple isothermal molecular method called Real-Time Fluorescence Loop-Mediated Isothermal Amplification (RealAmp) for the diagnosis of malaria. Currently, RealAmp primers exist for detecting the Plasmodium genus and the detection of P. falciparum and P. vivax species. The RealAmp method has great potential as a molecular tool for the diagnosis of malaria in the field (and other infections of major public health impact, such as HIV and tuberculosis). It can provide an alternative to conventional PCR-based diagnostic methods for field use in clinical and operational programs. The objective of this proposal is to validate the sensitivity of RealAmp for detection of malaria parasites in blood spots from patients with clinical diagnosis of malaria in two endemic states of Brazil with co-circulation of P. falciparum and P. vivax. In this evaluation, RealAmp and microscopic examination will be compared to a real-time PCR method as a reference test.

Condition or Disease	Intervention/Treatment	Phase
Malaria Diagnosis	Other: Realtime amplification (RealAmp)

Study Design

• Study Type: Observational
• Actual Enrollment: 1000 participants
• Observational Model: Cohort
• Time Perspective: Cross-Sectional
• Official Title: Validation of RealAmp Method for the Diagnosis of Malaria in Endemic Areas of Brazil
• Actual Study Start Date: Jan 1, 2015
• Actual Primary Completion Date: Jun 1, 2015
• Actual Study Completion Date: Jun 1, 2015

Outcome Measures

Primary Outcome Measures

1. Malaria Diagnosis Using RealAmp and Microscopy. [12 months]

   These are the results of the different tests used in the evaluation.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 7 Years and Older
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No

Inclusion Criteria:

• Age 7 years

• Signed informed consent for patients 18 years and older

• Signed parental/guardian consent form and assent from the patient (for those less than 18 years)

• Documented fever (auxiliary temperature greater than 37.5C) or self-reported fever in previous 24 hours in the absence of another obvious cause of fever

• First visit for current illness at the diagnostic post

Exclusion Criteria:

• Absence of fever or no history of fever

• Follow-up visit for the current illness

• No consent/assent to participate

• Antimalarial treatment use in previous 30 days

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Hospital do Jurua	Cruzeiro do Sul		Brazil

Sponsors and Collaborators

• Centers for Disease Control and Prevention
• Evandro Chagas Institute of Clinical Research

Investigators

• Principal Investigator: Alexandre Macedo de Oliveira, MD, Centers for Disease Control and Prevention

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

• Responsible Party: Centers for Disease Control and Prevention
• ClinicalTrials.gov Identifier: NCT02371395
• Other Study ID Numbers: 6619
• First Posted: Feb 25, 2015
• Last Update Posted: Oct 23, 2018
• Last Verified: May 1, 2015

• Additional relevant MeSH terms: Malaria; Disease

Study Results

Participant Flow

Arm/Group Title	Patients With Suspected Malaria (Fever)
Arm/Group Description	This was a study of enrolling patients with fever or history of fever for malaria testing. A total of 1,000 samples were collected from patients in Cruzeiro do Sul, Acre state from January to June, 2015.
Period Title: Overall Study
STARTED	1000
COMPLETED	1000
NOT COMPLETED	0

Baseline Characteristics

Arm/Group Title	Single Arm
Arm/Group Description	This was a study of enrolling patients with fever or history of fever for malaria testing. A total of 1,000 samples were collected from patients in Cruzeiro do Sul, Acre state from January to June, 2015.
Overall Participants	1000
Age (years) [Median (Full Range) ]
Median (Full Range) [years]	29.5
Sex: Female, Male (Count of Participants)
Female	551 55.1%
Male	449 44.9%
Race and Ethnicity Not Collected (Count of Participants)
Region of Enrollment (Count of Participants)
Brazil	1000 100%

Outcome Measures

1. Primary Outcome

Title	Malaria Diagnosis Using RealAmp and Microscopy.
Description	These are the results of the different tests used in the evaluation.
Time Frame	12 months

Outcome Measure Data

Analysis Population Description

Those were patients presenting with suspected malaria.

Arm/Group Title	Single Arm
Arm/Group Description	This was a study of enrolling patients with fever or history of fever for malaria testing. A total of 1,000 samples were collected from patients in Cruzeiro do Sul, Acre state from January to June, 2015.
Measure Participants	1000
P. falciparum	42 4.2%
P. vivax	153 15.3%
Mixed infection	0 0%
Species unidentified	1 0.1%
Negative	804 80.4%
P. falciparum	56 5.6%
P. vivax	169 16.9%
Mixed infection	1 0.1%
Species unidentified	0 0%
Negative	774 77.4%
P. falciparum	55 5.5%
P. vivax	181 18.1%
Mixed infection	0 0%
Species unidentified	3 0.3%
Negative	761 76.1%
P. falciparum	48 4.8%
P. vivax	152 15.2%
Mixed infection	20 2%
Species unidentified	0 0%
Negative	780 78%

Adverse Events

Arm/Group Title	Single Arm
Arm/Group Description	This was a study of enrolling patients with fever or history of fever for malaria testing. A total of 1,000 samples were collected from patients in Cruzeiro do Sul, Acre state from January to June, 2015.
All Cause Mortality
	Single Arm
	Affected / at Risk (%)	# Events
Total	/ (NaN)
Serious Adverse Events
	Single Arm
	Affected / at Risk (%)	# Events
Total	0/1000 (0%)
Other (Not Including Serious) Adverse Events
	Single Arm
	Affected / at Risk (%)	# Events
Total	0/1000 (0%)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.