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Filariasis IgG/IgM Rapid Test Device
Package Insert
Specimens: Whole Blood/Serum/Plasma
Effective Date: 2015‐02
For professional in vitro diagnostic use only.
INTENDED USE
The Filariasis IgG/IgM Rapid Test is a lateral flow immunoassay for the simultaneous detection and differentiation of IgG and IgM anti‐lymphatic filarial parasites (W. Bancrofti and B. Malayi) in human serum, plasma or whole blood. This test is intended to be used as a screening test and as an aid in
the diagnosis of infection with lymphatic filarial parasites. Any reactive specimen with the Filariasis IgG/IgM Rapid Test must be confirmed with alternative testing method(s).
The lymphatic filariasis known as Elephantiasis, mainly caused by W. bancrofti and B. malayi, affects about 120 million people over 80 countries1,2. The disease is transmitted to humans by the bites of infected mosquitoes within which the microflariae sucked from an infected human subject develops
into third‐stage larvae. Generally, repeated and prolonged exposure to infected larvae is required for establishment of human infection.
The definitive parasitologic diagnosis is the demonstration of microflariae in blood samples3. However, this gold standard test is restricted by the requirement for nocturnal blood collection and
lack of adequate sensitivity. Detection of circulating antigens is commercially available. Its usefulness is limited for W. bancrofti4. In addition, microfilaremia and antigenemia develop from months to years after exposure.
Antibody detection provides an early means to detect filarial parasite infection. Presence of IgM to the parasite antigens suggest current infection, whereas, IgG corresponds to late stage of infection or past infection5. Furthermore, identification of conserved antigens allows ‘pan‐filaria’ test to be applicable. Utilization of recombinant proteins eliminates cross‐reaction with individuals having
other parasitic diseases6. The Filariasis IgG/IgM Rapid Test uses conserved recombinant antigens to simultaneously detect IgG and IgM to the W. bancrofti and B. malayi parasites without the restriction on specimen collection.
The Filariasis IgG/IgM Rapid Test is a lateral flow chromatographic immunoassay. The test cassette consists of: 1) a burgundy colored conjugate pad containing recombinant W. bancrofti and B. malayi common antigens conjugated with colloid gold (Filariasis conjugates) and rabbit IgG‐gold conjugates, 2) a nitrocellulose membrane strip containing two test bands (T1 and T2 bands) and a control band (C band). The T1 band is pre‐coated with monoclonal anti‐human IgM for the detection of IgM anti‐ W. bancrofti and B. malayi, T2 band is pre‐coated with reagents for the detection of IgG anti‐ W. bancrofti and B. malayi, and the C band is pre‐coated with goat anti rabbit IgG.
When an adequate volume of test specimen is dispensed into the sample well of the cassette, the specimen migrates by capillary action across the cassette. W. bancrofti or B. malayi IgM antibodies if present in the specimen will bind to the Filariasis conjugates. The immunocomplex is then captured on the membrane by the pre‐coated anti‐human IgM antibody, forming a burgundy colored T2 band, indicating a W. bancrofti or B. malayi IgM positive test result.
W. bancrofti or B. malayi IgG antibodies if present in the specimen will bind to the Filariasis conjugates. The immunocomplex is then captured by the pre‐coated reagents on the membrane, forming a burgundy colored T1 band, indicating a W. bancrofti or B. malayi IgG positive test result. Absence of any test bands (T1 and T2) suggests a negative result. The test contains an internal control (C band) which should exhibit a burgundy colored band of the immunocomplex of goat anti rabbit IgG/rabbit IgG‐gold conjugate regardless of the color development on any of the test bands. Otherwise, the test result is invalid and the specimen must be retested with another device.
Each device contains a strip with colored conjugates
Individually packed test devices and reactive reagents pre‐spreaded at the
corresponding regions
Disposable pipettes For adding specimens use
Buffer Phosphate buffered saline and preservative
Package insert For operation instruction
STORAGE AND STABILITY
Store as packaged in the sealed pouch either at room temperature or refrigerated (2‐30°C). The test device is stable through the expiration date printed on the sealed pouch. The test device must remain in the sealed pouch until use. Do not freeze. Do not use beyond the expiration date.
Materials Provided
Materials Required But Not Provided
|
|
For professional in vitro diagnostic use only.
Do not use after expiration date indicated on the package. Do not use the test if its foil pouch is
damaged. Do not reuse tests.
This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary
state of the animals does not totally guarantee the absence of transmissible pathogenic agents. It is therefore, recommended that these products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest or inhale).
Avoid cross‐contamination of specimens by using a new specimen collection container for each
specimen obtained.
Read the entire procedure carefully prior to performing any tests.
Do not eat, drink or smoke in the area where the specimens and kits are handled. Handle all
specimens as if they contain infectious agents. Observe established precautions against microbiological hazards throughout the procedure and follow the standard procedures for proper disposal of specimens. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimens are assayed.
Do not interchange or mix reagents from different lots.
Humidity and temperature can adversely affect results.
SPECIMEN COLLECTION AND HANDLING
1. The Filariasis IgG/IgM Rapid Test can be performed used on Whole Blood /Serum / Plasma.
2. To collect whole blood, serum or plasma specimens following regular clinical laboratory procedures.
3. Testing should be performed immediately after specimen collection. Do not leave the specimens at room temperature for prolonged periods. For long term storage, specimens should be kept below ‐ 20C. Whole blood should be stored at 2‐8C if the test is to be run within 2 days of collection. Do not freeze whole blood specimens.
4. Bring specimens to room temperature prior to testing. Frozen specimens must be completely thawed and mixed well prior to testing. Specimens should not be frozen and thawed repeatedly.
ASSAY PROCEDURE
1. Bring the specimen and test components to room temperature. Place the test device on a clean, flat surface.
2. Fill the pipette dropper with the specimen. Apply 1 drop of specimen (whole blood, plasma or serum) into the sample well, then add 1 drop (about 35‐50 µL) of Sample Diluent immediately. Set up timer.
3. Results can be read in 15 minutes. Don’t read result after 15 minutes.
INTERPRETATION OF ASSAY RESULT
POSITIVE RESULT:
C
T1
T2
C
T1
T2
T1
T2
*NOTE: The intensity of the color in the test line region(s) will vary depending on the concentration of Filariasis antibodies in the specimen. Therefore, any shade of color in the test line region(s) should be considered positive.
NEGATIVE RESULT:
C
T1
T2
INVALID C T1 T2 | RESULT: C T1 T2 | Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test device. If the problem persists, discontinue using the test kit immediately and contact your local distributor. |
Using individual Filariasis IgG/IgM Rapid Test cassettes as described in the Assay Procedure above, run 1 Positive Control and 1 Negative Control (provided upon request) under the following circumstances to monitor test performance:
1. A new operator uses the kit, prior to performing testing of specimens.
2. A new test kit is used.
3. A new shipment of kits is used.
4. The temperature used during storage of the kit falls outside of 2°C‐30°C.
5. The temperature of the test area falls outside of 15°C‐30°C.
1. Clinical Performance For IgM Test
24 samples from patients with acute lymphatic filariasis and 200 samples collected from a non‐filariasis region were tested by the Filariasis IgG/IgM Rapid Test.
Comparison for all subjects is showed in the following table:
Filariasis IgG/IgM Rapid Test | |||
Clinical Status | Positive | Negative | Total |
Acute filariasis | 23 | 1 | 24 |
Negative | 0 | 200 | 200 |
Total | 23 | 201 | 224 |
Relative Sensitivity: 95.8%; Relative Specificity: 100%; Overall agreement: 99.6%
2. Clinical Performance For IgG Test
26 samples from patients with chronic lymphatic filariasis and 200 samples collected from a non‐filariasis region were tested by the Filariasis IgG/IgM Rapid Test. Comparison for all subjects is showed in the following table:
Filariasis IgG/IgM Rapid Test | |||
Clinical Status | Positive | Negative | Total |
Chronic filariasis | 24 | 2 | 26 |
Negative | 0 | 200 | 200 |
Total | 24 | 202 | 226 |
Relative Sensitivity: 92.3%; Relative Specificity: 100%; Overall agreement: 99. 1%
1. The Assay Procedure and the Test Result Interpretation must be followed closely when testing the presence of antibodies to filarial parasites in serum, plasma or whole blood from individual subjects. Failure to follow the procedure may give inaccurate results.
2. The Filariasis IgG/IgM Rapid Test is limited to the qualitative detection of antibodies to W. bancrofti and B. malayi in human serum, plasma or whole blood. The intensity of the test band does not have linear correlation with the antibody titer in the specimen.
3. A negative result for an individual subject indicates absence of detectable W. bancrofti and B. malayi antibodies. However, a negative test result does not preclude the possibility of exposure to W. bancrofti and B. malayi.
4. A negative result can occur if the quantity of W. bancrofti and B. malayi antibodies present in the specimen is below the detection limits of the assay, or the antibodies that are detected are not present during the stage of disease in which a sample is collected.
5. Some specimens containing unusually high titer of heterophile antibodies or rheumatoid factor may affect expected results.
6. The results obtained with this test should only be interpreted in conjunction with other diagnostic procedures and clinical findings.
1. Lymphatic filariasis: the disease and its control. Fifth report of the WHO Expert Committee on Filariasis. WHO Tech Rep Ser 1992; 281‐871.
2. Michael E, Bundy DAP, Grenfell BT. Re‐assessing the global prevalence and distribution of lymphatic filariasis. Parasitology 1996; 112:405‐428.
3. Eberhard ML, Lammie PJ. Laboratory diagnosis of filariasis. Clin. Lab Med 1991; 11:977‐ 1010.
4. More SJ, Copeman DB. A highly specific and sensitive monoclonal antibody‐based ELISA for the detection of circulating antigen in bancroftian filariasis. Trop Med Parasitol 1990; 41:403‐406
5. Lammie PJ, Weil G, et al: Recombinant antigen‐based antibody assays for the diagnosis surveillance of lymphatic filariasis‐a multiplecenter trial. Flaria Jornal 2004: 3: 9‐ 18.
Filariasis IgG/IgM Rapid Test Device
Package Insert
Specimens: Whole Blood/Serum/Plasma
Effective Date: 2015‐02
For professional in vitro diagnostic use only.
INTENDED USE
The Filariasis IgG/IgM Rapid Test is a lateral flow immunoassay for the simultaneous detection and differentiation of IgG and IgM anti‐lymphatic filarial parasites (W. Bancrofti and B. Malayi) in human serum, plasma or whole blood. This test is intended to be used as a screening test and as an aid in
the diagnosis of infection with lymphatic filarial parasites. Any reactive specimen with the Filariasis IgG/IgM Rapid Test must be confirmed with alternative testing method(s).
The lymphatic filariasis known as Elephantiasis, mainly caused by W. bancrofti and B. malayi, affects about 120 million people over 80 countries1,2. The disease is transmitted to humans by the bites of infected mosquitoes within which the microflariae sucked from an infected human subject develops
into third‐stage larvae. Generally, repeated and prolonged exposure to infected larvae is required for establishment of human infection.
The definitive parasitologic diagnosis is the demonstration of microflariae in blood samples3. However, this gold standard test is restricted by the requirement for nocturnal blood collection and
lack of adequate sensitivity. Detection of circulating antigens is commercially available. Its usefulness is limited for W. bancrofti4. In addition, microfilaremia and antigenemia develop from months to years after exposure.
Antibody detection provides an early means to detect filarial parasite infection. Presence of IgM to the parasite antigens suggest current infection, whereas, IgG corresponds to late stage of infection or past infection5. Furthermore, identification of conserved antigens allows ‘pan‐filaria’ test to be applicable. Utilization of recombinant proteins eliminates cross‐reaction with individuals having
other parasitic diseases6. The Filariasis IgG/IgM Rapid Test uses conserved recombinant antigens to simultaneously detect IgG and IgM to the W. bancrofti and B. malayi parasites without the restriction on specimen collection.
The Filariasis IgG/IgM Rapid Test is a lateral flow chromatographic immunoassay. The test cassette consists of: 1) a burgundy colored conjugate pad containing recombinant W. bancrofti and B. malayi common antigens conjugated with colloid gold (Filariasis conjugates) and rabbit IgG‐gold conjugates, 2) a nitrocellulose membrane strip containing two test bands (T1 and T2 bands) and a control band (C band). The T1 band is pre‐coated with monoclonal anti‐human IgM for the detection of IgM anti‐ W. bancrofti and B. malayi, T2 band is pre‐coated with reagents for the detection of IgG anti‐ W. bancrofti and B. malayi, and the C band is pre‐coated with goat anti rabbit IgG.
When an adequate volume of test specimen is dispensed into the sample well of the cassette, the specimen migrates by capillary action across the cassette. W. bancrofti or B. malayi IgM antibodies if present in the specimen will bind to the Filariasis conjugates. The immunocomplex is then captured on the membrane by the pre‐coated anti‐human IgM antibody, forming a burgundy colored T2 band, indicating a W. bancrofti or B. malayi IgM positive test result.
W. bancrofti or B. malayi IgG antibodies if present in the specimen will bind to the Filariasis conjugates. The immunocomplex is then captured by the pre‐coated reagents on the membrane, forming a burgundy colored T1 band, indicating a W. bancrofti or B. malayi IgG positive test result. Absence of any test bands (T1 and T2) suggests a negative result. The test contains an internal control (C band) which should exhibit a burgundy colored band of the immunocomplex of goat anti rabbit IgG/rabbit IgG‐gold conjugate regardless of the color development on any of the test bands. Otherwise, the test result is invalid and the specimen must be retested with another device.
Each device contains a strip with colored conjugates
Individually packed test devices and reactive reagents pre‐spreaded at the
corresponding regions
Disposable pipettes For adding specimens use
Buffer Phosphate buffered saline and preservative
Package insert For operation instruction
STORAGE AND STABILITY
Store as packaged in the sealed pouch either at room temperature or refrigerated (2‐30°C). The test device is stable through the expiration date printed on the sealed pouch. The test device must remain in the sealed pouch until use. Do not freeze. Do not use beyond the expiration date.
Materials Provided
Materials Required But Not Provided
|
|
For professional in vitro diagnostic use only.
Do not use after expiration date indicated on the package. Do not use the test if its foil pouch is
damaged. Do not reuse tests.
This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary
state of the animals does not totally guarantee the absence of transmissible pathogenic agents. It is therefore, recommended that these products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest or inhale).
Avoid cross‐contamination of specimens by using a new specimen collection container for each
specimen obtained.
Read the entire procedure carefully prior to performing any tests.
Do not eat, drink or smoke in the area where the specimens and kits are handled. Handle all
specimens as if they contain infectious agents. Observe established precautions against microbiological hazards throughout the procedure and follow the standard procedures for proper disposal of specimens. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimens are assayed.
Do not interchange or mix reagents from different lots.
Humidity and temperature can adversely affect results.
SPECIMEN COLLECTION AND HANDLING
1. The Filariasis IgG/IgM Rapid Test can be performed used on Whole Blood /Serum / Plasma.
2. To collect whole blood, serum or plasma specimens following regular clinical laboratory procedures.
3. Testing should be performed immediately after specimen collection. Do not leave the specimens at room temperature for prolonged periods. For long term storage, specimens should be kept below ‐ 20C. Whole blood should be stored at 2‐8C if the test is to be run within 2 days of collection. Do not freeze whole blood specimens.
4. Bring specimens to room temperature prior to testing. Frozen specimens must be completely thawed and mixed well prior to testing. Specimens should not be frozen and thawed repeatedly.
ASSAY PROCEDURE
1. Bring the specimen and test components to room temperature. Place the test device on a clean, flat surface.
2. Fill the pipette dropper with the specimen. Apply 1 drop of specimen (whole blood, plasma or serum) into the sample well, then add 1 drop (about 35‐50 µL) of Sample Diluent immediately. Set up timer.
3. Results can be read in 15 minutes. Don’t read result after 15 minutes.
INTERPRETATION OF ASSAY RESULT
POSITIVE RESULT:
C
T1
T2
C
T1
T2
T1
T2
*NOTE: The intensity of the color in the test line region(s) will vary depending on the concentration of Filariasis antibodies in the specimen. Therefore, any shade of color in the test line region(s) should be considered positive.
NEGATIVE RESULT:
C
T1
T2
INVALID C T1 T2 | RESULT: C T1 T2 | Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test device. If the problem persists, discontinue using the test kit immediately and contact your local distributor. |
Using individual Filariasis IgG/IgM Rapid Test cassettes as described in the Assay Procedure above, run 1 Positive Control and 1 Negative Control (provided upon request) under the following circumstances to monitor test performance:
1. A new operator uses the kit, prior to performing testing of specimens.
2. A new test kit is used.
3. A new shipment of kits is used.
4. The temperature used during storage of the kit falls outside of 2°C‐30°C.
5. The temperature of the test area falls outside of 15°C‐30°C.
1. Clinical Performance For IgM Test
24 samples from patients with acute lymphatic filariasis and 200 samples collected from a non‐filariasis region were tested by the Filariasis IgG/IgM Rapid Test.
Comparison for all subjects is showed in the following table:
Filariasis IgG/IgM Rapid Test | |||
Clinical Status | Positive | Negative | Total |
Acute filariasis | 23 | 1 | 24 |
Negative | 0 | 200 | 200 |
Total | 23 | 201 | 224 |
Relative Sensitivity: 95.8%; Relative Specificity: 100%; Overall agreement: 99.6%
2. Clinical Performance For IgG Test
26 samples from patients with chronic lymphatic filariasis and 200 samples collected from a non‐filariasis region were tested by the Filariasis IgG/IgM Rapid Test. Comparison for all subjects is showed in the following table:
Filariasis IgG/IgM Rapid Test | |||
Clinical Status | Positive | Negative | Total |
Chronic filariasis | 24 | 2 | 26 |
Negative | 0 | 200 | 200 |
Total | 24 | 202 | 226 |
Relative Sensitivity: 92.3%; Relative Specificity: 100%; Overall agreement: 99. 1%
1. The Assay Procedure and the Test Result Interpretation must be followed closely when testing the presence of antibodies to filarial parasites in serum, plasma or whole blood from individual subjects. Failure to follow the procedure may give inaccurate results.
2. The Filariasis IgG/IgM Rapid Test is limited to the qualitative detection of antibodies to W. bancrofti and B. malayi in human serum, plasma or whole blood. The intensity of the test band does not have linear correlation with the antibody titer in the specimen.
3. A negative result for an individual subject indicates absence of detectable W. bancrofti and B. malayi antibodies. However, a negative test result does not preclude the possibility of exposure to W. bancrofti and B. malayi.
4. A negative result can occur if the quantity of W. bancrofti and B. malayi antibodies present in the specimen is below the detection limits of the assay, or the antibodies that are detected are not present during the stage of disease in which a sample is collected.
5. Some specimens containing unusually high titer of heterophile antibodies or rheumatoid factor may affect expected results.
6. The results obtained with this test should only be interpreted in conjunction with other diagnostic procedures and clinical findings.
1. Lymphatic filariasis: the disease and its control. Fifth report of the WHO Expert Committee on Filariasis. WHO Tech Rep Ser 1992; 281‐871.
2. Michael E, Bundy DAP, Grenfell BT. Re‐assessing the global prevalence and distribution of lymphatic filariasis. Parasitology 1996; 112:405‐428.
3. Eberhard ML, Lammie PJ. Laboratory diagnosis of filariasis. Clin. Lab Med 1991; 11:977‐ 1010.
4. More SJ, Copeman DB. A highly specific and sensitive monoclonal antibody‐based ELISA for the detection of circulating antigen in bancroftian filariasis. Trop Med Parasitol 1990; 41:403‐406
5. Lammie PJ, Weil G, et al: Recombinant antigen‐based antibody assays for the diagnosis surveillance of lymphatic filariasis‐a multiplecenter trial. Flaria Jornal 2004: 3: 9‐ 18.
Test devices
Buffer
Specimen collection containers
Centrifuge
Droppers
Package insert
Heparinized capillary tubes 
Timer