Microsoft word - ddcr# 13-384 900121-03 ce cl detect rapid test package insert

Precautions
CL Detect™ Rapid Test
 Adhere to the procedures described in this insert. Any For Cutaneous Leishmaniasis
variations from the described methodology may affect For Export Use Only
 Use this test only with skin lesion samples. Do not use The CL Detect™ Rapid Test is a qualitative, in vitro serum, plasma or whole blood with this test strip. immunochromatographic assay for the rapid detection of  Do not use samples from lesions that are over 2 months Leishmania species in skin lesions from an individual old or samples that have undergone multiple freeze- suspected of having cutaneous leishmaniasis. This test has been designed to test for species of the genus Leishmania  Handle all samples and kits used as if they contain that cause cutaneous leishmaniasis (CL). This test is infectious agents. Observe established precautions intended for laboratory professional use only and is for against microbiological hazards while performing all procedures and follow the standard procedures for proper disposal of samples and used kits. Summary and Explanation
 Wear protective clothing, eye protection and disposable gloves while performing the assay. Wash hands Several Leishmania species are responsible for cutaneous leishmaniasis. In the Middle East and Central Asia, the  Avoid all contact between hands and eyes or other predominant species responsible for cutaneous forms of leishmaniasis are L. major and L. tropica. L. donovani and  Do not eat, drink or smoke in the area where the L. infantum predominantly lead to visceral forms of Leishmaniasis. In Iraq, L. major is the major cause of  Chase Buffer contains a preservative; avoid all possible cutaneous leishmaniasis with most of the reported cases in contact with skin and mucous membranes. Lysis buffer the army being due to this agent. It is the primary agent for contains detergents and should not be in contact with zoonotic cutaneous leishmaniasis (ZCL). In Afghanistan, the primary agent for cutaneous leishmaniasis is L. tropica with an active infection rate in Kabul of 2.7%, but areas of Northern Afghanistan are also endemic for L. major. L. tropica is more frequently associated with anthroponotic cutaneous leishmaniasis (ACL). In Central and South The entire kit is designed to be stored at room temperature America, L. braziliensis, mexicana, amazonensis and (20C-30C) for the duration of its shelf life. Exposure to panamensis, among other species, are prevalent and temperatures over 30C can impact the performance of the responsible for cutaneous leishmaniasis. test and should be minimized. The test strips should not be frozen. The test strips should be used immediately after Principle
removal from the pouch or vial to prevent exposure to The CL Detect™ Rapid Test for CL is a qualitative, membrane based immunoassay for the detection of Kit Contents
amastigote antigens present in skin lesions of individuals infected with Leishmania parasites. The membrane is pre- CL DetectTM Rapid Test strip's membrane is pre-coated with coated with an affinity purified polyclonal antibody to an an affinity purified polyclonal antibody that reacts with amastigote antigen (Peroxidoxin) on the test line region and leishmania amastigotes on the test line region and goat anti goat anti-mouse IgG on the control line region. During mouse IgG on the control line region. It is detected with a testing, the lysed sample reacts with the dye conjugate colloidal gold labeled monoclonal antibody to the same (monoclonal antibody to the amastigote antigen) which has been pre-coated in the test device. The mixture then chromatographically migrates upward on the membrane, The kit contains the following: reacts with the affinity purified rabbit antibody, generating a 1. Twenty-five (25) individually pouched test strips or red line. Presence of this red line indicates a positive result, twenty-five (25) test strips in a vial with desiccant in the while its absence indicates a negative result. Regardless of the presence of amastigote antigens, as the mixture continues to migrate across the membrane to the immobilized goat 3. One (1) vial of Chase Buffer solution, 6ml. anti-mouse IgG region, a red line at the control line region 4. One (1) vial of Positive Control solution, 6ml. will always appear. The presence of this red line serves as 5. One (1) vial of Negative Control solution, 6ml. verification for sufficient sample volume and proper flow CE CL Detect Rapid Test Insert Part No. 900121-03
Effective Date: 07/08/2013
The following materials are provided optionally:  Press a thumb on to the skin where the broach was 2. Twenty-five (25) sterile dental broaches.  Gently rotate (twist) the broach twice (back and 3. Twenty-five (25) transfer pipets (20ul).  Remove broach with a quick sharp pull, twisting Materials needed but not provided:  Place it barbed-end down into a sample cup  Twist the barb to remove as much cellular material Sample Collection
 Use transfer pipette to flush tissue from broach as  Only skin lesions are to be used with this kit. Human serum should not be tested with this kit.  Make sure the lysis buffer and material are well  Samples must be treated with 3 drops of lysis buffer in mixed. Material should remain in lysis buffer for 5- preparation for testing. Incubate samples in lysis buffer 10 minutes prior to testing with CL Detect™. at room temperature for 5-10 minutes. Samples must not remain in lysis buffer for more than 30 minutes prior to Dermal Scraping Procedure
testing. In all cases, ensure that the lysis buffer and the  Unwrap sterile scalpel blade/sterile vaccinostyle from packaging. Note that this is not provided with  Test should be performed as soon as possible after collection of the sample. Do not leave samples at room  Lift the necrotic lip of the lesion to reach the area temperature for prolonged periods. Samples can be refrigerated at 2-8C up to 1 day. Otherwise samples  Scrape the upper dermis 4 times from the inside in an outward motion using the edge of the scalpel  Bring sample to room temperature prior to testing. Frozen samples must be completely thawed prior to  Deposit tissue flecks and juice directly from the testing. Samples should not be repeatedly frozen and surface of the blade into a collection cup (may be provided with this kit) by rinsing with 3 drops of  If samples are to be shipped, they should be packed in compliance with Federal Regulations covering  Make sure the lysis buffer and material are well mixed. Material should remain in lysis buffer for 5- 10 minutes prior to testing with CL Detect™. Lesion Preparation: A sample should be obtained by a
trained medical professional. Note that the lesion should
Test Procedure
be less than 2 months old. Guidelines for preparing the
lesion to obtain a sample, is described as follows:
Refer to “Test Procedure” figure on page 3.
 Thoroughly clean the lesion and surrounding skin 1. Collect skin lesion sample as described in the Sample  Debride the sore by using forceps to apply a gauze Collection section of this product insert. pad soaked in sterile 0.9% saline solution. Rub until 2. Allow the sample to reach room temperature prior to the entire crust is removed, leaving the lesion testing. Sample should remain in lysis buffer for 5-10 minutes prior to testing with CL Detect™.  Apply EMLA cream to the surrounding edge of the 3. Remove the test strip from the foil pouch or vial. lesion or use injectable lidocaine if necessary to 4. Add 20 l of sample to the test strip in the area beneath  A scalpel may be used to debride if necessary, only 5. Add 3 drops of the Chase Buffer solution provided with after reducing discomfort pain with EMLA or 6. Place the test strip into the cup so that the end of the  Once the lesion is prepared, a sample can be strip is facing downward as indicated by the arrows on obtained by a trained medical professional. Below are two methods that have been clinically proven to 7. Read the results in 20-30 minutes. It is important that the background is clear before reading the test, especially when samples have low levels of antigen, and Dental Broach Procedure
only a weak band appears in the test region (T). Results  Remove dental broach from individual packaging. interpreted after 30 minutes can be misleading.  At the border of the lesion, insert the barb into the lesion to a depth of approximately ½ the length of Note: Do not test this product with the Chase Buffer
the barb from the ulcer to the inflamed area. solution alone. 20 l of sample must be added first. CE CL Detect Rapid Test Insert Part No. 900121-03
Effective Date: 07/08/2013
Controls
Interpretation of Results
One vial each of positive and negative control is provided Refer to “Interpretation” figure on page 3.
with each test kit. Positive and negative controls should be tested with each new kit of 25 test strips to ensure kit A Positive Result
integrity. Avoid switching caps of positive and negative The test is positive when a control line and test line appear
control bottles, or otherwise contaminating the tips.
in the test area. A positive result indicates that the CL DetectTM Rapid Test detected antigens present in Procedure:
amastigotes. A faint line is considered a positive result. As 1. Dilute positive or negative sample by adding 1 drop of a guide for interpretation, the red color in the test region will positive or negative control plus 3 drops of lysis buffer vary depending on the concentration of antigens present. The test line for "weakly positive" samples may show results 2. Remove test strip from pouch or vial. between a weak positive red line to a faintly red, almost 3. Add 20 l of diluted positive or negative control to the white background. "Weakly positive" samples are those with test strip in the area beneath the arrow using a transfer 4. Add 3 drops of Chase Buffer solution to a reaction cup. A Negative Result
5. Place the test strip into the cup so that the end of the The test is negative when only the control line appears. A strip is facing downward as indicated by the arrows on negative result indicates that the CL Detect™ Rapid Test did not detect antigens in the sample. No test line is present. 6. Read the results in 20-30 minutes. Follow procedure below regarding interpretation of results. An Invalid Result
7. The positive control should yield a positive result, and No control line is visible. It is recommended to retest using a new CL Detect™ Rapid Test for Cutaneous Leishmaniasis 8. If either the positive or negative control fails, retest with Note: The red color in the test region will vary depending
on the concentration of Leishmanial antigen present. However, neither the quantitative value nor the rate of increase in antigen can be determined by this qualitative test. CE CL Detect Rapid Test Insert Part No. 900121-03
Effective Date: 07/08/2013
Performance Characteristics
considered with other clinical information available to  If the result is negative and clinical symptoms persist, In endemic areas, the sensitivity of the CL Detect™ Rapid additional follow-up testing using other clinical methods Test is to be determined. The specificity of the test may is recommended. A negative result does not preclude  A false positive result may occur. Confirmatory testing Comparison of CL Detect™ Rapid Test to Microscopy
(such as by culture) is advised especially in cases where for Dental Broach collection method
Fifty-nine (59) Cutaneous leishmaniasis skin lesion samples were collected using a dental broach collection device under  The performance of this test has not been evaluated with the IRB-approved protocol for study sites in Tunisia. Samples, typically from lesions <2 months old, were  Use of bloody samples may lead to inaccurate results. collected in lysis buffer and tested in the CL Detect™ Rapid Residual betadine or mercurochrome on samples may Test. The CL Detect™ Rapid Test reactivity was compared lead to inaccurate results. Lesions must be thoroughly to microscopy data (WHO scale) for the same lesion (see cleaned and debrided as directed in the “Sample Collection” section of this insert, prior to sample Microscopy (WHO)
References
CL DetectTM
1. Martin SK, Thuita-Harun L, Adoyo-Adoyo M, (with Dental Broach)
0 4 Wasunna KM. 1998. A diagnostic ELISA for visceral leishmaniasis, based on antigen from media conditioned by Leishmania donovani promastigotes. Ann Trop Med In a separate study, samples from skin lesions that are not due to Leishmaniasis were collected using the dental broach. 2. Ryan JR, Smithyman AM,Stiteler JM, Rowton ED, To date, 16 samples with non-CL skin lesions have been Valli L, Orndorff GR, Seaman J, Thuita-Harun L and tested using the Dental Broach collection method. All were Martin SK. An ELISA based on soluble promastigote negative by CL Detect™ Rapid Test and Microscopy for antigen detects IgM and IgG antibodies in visceral and cutaneous leishmaniasis. Abstract to November, 1998 American Society for Tropical Medicine and Hygiene Microscopy (WHO)
3. Ryan JR, Rajasekariah GH, Thuita-Harun L, Yi L, Smithyman AM, Kinnamon KE, and Martin SK. The CL DetectTM
0 0 potential use of antigens shed, excreted and secreted by (with Dental Broach)
0 16 Leishmania donovani promastigotes for antigen- detection tests for leishmaniasis. Abstract to December, 1999 American Society for Tropical Medicine and Performance characteristics for CL Detect™ Rapid Test in 4. Adoyo MA,Magiri CG,Ryan JR, Martin SK, Mason CJ and Wasunna KM. Preliminary applications of an antibody capture ELISA to monitor leishmaniasis therapy. Abstract to December, 1999 American Society for Tropical Medicine and Hygiene Meeting, 5. Mimori T, Matsumoto T, Calvopina MH, Gomez EA, Saya H, Katakura K, Nonaka S, Shamsuzzaman SM and Hashiguchi Y. Usefulness of sampling with a cotton swab for PCR diagnosis of cutaneous leishmaniasis in Limitations
the New World. Acta Trop 2002; 81(3): 197-202. 6. Marques MJ, Volpini AC, Genari O, Mayrink W and  This test will only indicate the presence of Leishmania Romanha AJ. Simple form of clinical sample parasite antigens in patients with Cutaneous preservation and DNA extraction from human lesions Leishmaniasis and should not be used as the sole for the diagnosis of American cutaneous leishmaniasis criterion for the diagnosis of Leishmaniasis. This test via polymerase chain reaction. Am J Trop Med Hyg alone must not be used for any clinical treatment decision. As with all diagnostic tests, all results must be CE CL Detect Rapid Test Insert Part No. 900121-03
Effective Date: 07/08/2013
7. Mallik MK, Das DK and Haji BE. Fine needle 21. Sharquie KE, Hassen AS, Hassan SA, Al-Hamami IA. aspiration cytology diagnosis of cutaneous Evaluation of diagnosis of cutaneous leishmaniasis by leishmaniasis in a 6-month-old child: a case report. Acta direct smear, culture and histopathology. Saudi Med J 8. Ramirez JR, Aguledo S, Muskus C, Alzate JF, 22. Navin TR, Arana FE, deMerida AM, Arana BA, Berberich C, Barker D and Velez ID. Diagnosis of Castillo AL and Silvers DN. Cutaneous Leishmaniasis cutaneous leishmaniasis in Columbia: the sampling site in Guatemala: A comparison of Diagnostic methods. within lesions influences the sensitivity of parasitologic diagnosis. J Clin Microbiol. 2000; 38(10): 3768-73. 9. Matsumoto T, Hashiguchi Y, Gomez EA, Calvopina MH, Nonaka S, Saya H and Mimori T. Comparison of PCR results using scrape/exudates, syringe-sucked fluid and biopsy samples for diagnosis of cutaneous leishmaniasis in Ecuador. Trans R Soc Trop Med Hyg. 10. Mohareb EW, Hanafi HA, Mikhail EM, Presley SM and immunofluorescence assay for the detection of Leishmania promastigotes and amastigotes in sand flies and lesion fluid aspirates. J Egypt Soc Parasitol. 1998; 11. Webb JR, Campos-Neto A, Ovendale PJ, Martin TI, Stromberg EJ, Badaro R, Reed SG. Human and murine immune responses to a novel Leishmania major Immun. European Authorized Representative
12. Rafati S, Salmanian AH, Hashemi K, Schaff C, Belli S and Fasel N. Identification of Leishmania major cysteine proteases as targets of the immune response in humans. Mol Biochem Parasitol. 2001; 113(1) 35-43. 13. de Ibarra AA, Howard JG and Snary D. Monoclonal antibodies to Leishmania tropica major: specificities and antigen location. Parasitology 1982; 85(3): 523-31. 14. Greenblatt CL, Slutzky GM, de Ibarra AA and Snary D. Product is covered by the U.S. Patent # 8173383.
Monoclonal antibodies for serotyping Leishmania strains. J Clin Microbiol. 1983; 18(1): 191-3. 15. Handman E, Greenblatt CL and Goding JW. An ampipathic sulphated glycoconjugate of Leishmania: characterization with monoclonal antibodies. EMBO J 1984; 3(10): 2301-6. 16. Hailu A. The use of direct agglutination test (DAT) in serological diagnosis of Ethiopian cutaneous leishmaniasis. Diag Microbiol and Inf Dis 2002; 42: 251-256. 17. Beena KR, Ramesh V and Mukherjee A. Identification of parasite antigen, correlation of parasite density and inflammation in skin lesions of post kala-azar dermal leishmaniasis. J Cutan Pathol 2003; 30: 616-620. 18. Chargui N, Bastien P, Kallel K et al Usefulness of PCR in the diagnosis of cutaneous leishmaniasis in Tunisia. Trans Roy Soc Trop Med and Hyg 2005; 99, 762-768. 19. Robinson Ramirez J, Agudelo S, Muskus C et al Diagnosis of cutaneous leishmaniasis in Columbia: the sampling site within lesions influences the sensitivity of parasitologic diagnosis. J Clin Micro 2000; 38(10): 3768-3773. 20. Kassi M, Tareen I, Qazi A and Kasi PM. Fine needle aspiration cytology in the diagnosis of cutaneous leishmaniasis. Ann Saudi Med. 2004; 24(2): 93-97. CE CL Detect Rapid Test Insert Part No. 900121-03
Effective Date: 07/08/2013

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