Leishmania Antibody ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

Thi kit is intended for the qualitative determination of IgG class antibodies against Leishmania infantum in human serum or plasma (citrate, heparin).

Contents of Kit

1. Microtiterplate: 12 break-apart 8-well snap-off strips coated with Leishmania infantum antigens; in resealable aluminium foil.
2. IgG Sample Dilution Buffer: 1 bottle containing 100 mL of phosphate buffer (10 mM) for sample dilution; pH 7.2 ± 0.2; coloured yellow; ready to use; white cap; ≤ 0.0015% (v/v) CMIT/ MIT (3:1).
3. Stop Solution: 1 bottle containing 15 mL sulphuric acid, 0.2 mol/L; ready to use; red cap.
4. Washing Buffer (20× conc.): 1 bottle containing 50 mL of a 20-fold concentrated phosphate buffer (0.2 M), pH 7.2 ± 0.2, for washing the wells; white cap.
5. Conjugate: 1 bottle containing 20 mL of peroxidase Protein A in phosphate buffer (10 mM); coloured blue; ready to use; black cap.
6. TMB Substrate Solution: 1 bottle containing 15 mL 3,3',5,5'-tetramethylbenzidine (TMB), < 0.1 %; ready to use; yellow cap.
7. Positive Control: 1 vial containing 2 mL control; coloured yellow; ready to use; red cap; ≤ 0.02% (v/v) MIT.
8. Cut-off Control: 1 vial containing 3 mL control; coloured yellow; ready to use; green cap; ≤ 0.02% (v/v) MIT.
9. Negative Control: 1 vial containing 2 mL control; coloured yellow; ready to use; blue cap; ≤ 0.0015% (v/v) CMIT/ MIT (3:1).

Storage

Store the kit at 2-8 °C. The opened reagents are stable up to the expiry date stated on the label when stored at 2-8 °C.

Precision

General Description

Leishmania are protozoa belonging to the family trypanosomatidae. The parasites exist in two forms: the promastigotes in the midgut of the vector insect, and the amastigotes within the phagolysosomes of macrophages in their mammalian hosts. In the macrophages they live as round, non-motile amastigotes (3-7 μm in diameter). The macrophages are ingested by the sandfly during blood-meal and the amastigotes are released into their stomach. Almost immediately the amastigotes transform in to the motile, elongated (10-20μm), flagellate promastigote form, which migrate to the alimentary tract of the fly and after multiplication move forward to the salivary glands of the insect. Leishmaniases are a globally widespread group of parasitic diseases; the "type" is determined by the primary location of the macrophages that are infected. In humans four different forms of Leishmaniasis with a broad range of clinical manifestations are present; all can have devasting consequences. Leishmaniasis currently affects some 12 million people in 88 countries, all but 16 of which are in the developing world. It is estimated that 350 million people are exposed to the risk of infection by the different species of Leishmania parasite; the annual incidence of new cases is about 2 million (1-1.5 million cases of CL, 500,000 cases of VL). Visceral leishmaniasis (VL) is the most severe form of the disease, which, if untreated, has a mortality rate of almost 100%. Like many other tropical diseases, the leishmaniases are related to economic development and man-made environmental changes, which increase exposure to the sandfly vector. The geographical distribution is limited by the distribution of the sandfly. AIDS and other immunosuppressive conditions increase the risk of Leishmania infected people developing visceral illness (VL). Leishmania/HIV co-infections are considered to be a real "emerging disease", especially in southwestern Europe, where 25-70% of adult VL cases are related to HIV infection, and 1.5-9.5% of AIDS cases suffer from newly acquired or reactivated VL. Intravenous drug users have been identified as the main population at risk.