Specific antibodies in serum of patients with hydatidosis recognised by immunoblotting

Abstract

Summary. Hydatid fluids from sheep, goat, pig and man, after resolution by sodium dodecyl sulphate-polyacrylamide gel electrophoresis under reducing conditions, revealed at least 1 5 discrete polypeptide bands of 8-1 16 Kda. By ELISA, sera from all 20 cases of hydatidosis showed anti-hydatid antibody, but so did 11 (73%) of 15 sera samples from cysticercosis patients, eight (67%) of 12 sera from patients with other parasitic infections (amoebic liver abscess or hymenolepiasis) and one (4%) of 25 sera from healthy controls. Antibody to cysticercus antigen was found in 14 (93%) of 15 sera from cysticercosis patients, 17 (85%) of 20 sera from hydatid patients, six (50%) of 12 sera from patients with other parasitic infections and one (4%) of 25 sera from healthy controls. Sera from 17 (85%) of 20 hydatid patients, 11 (73%) of 15 cysticercosis patients and five (42%) of 12 patients with other parasitic infections had antibodies to both hydatid and cysticercus antigens. Sera from 20 surgically confirmed cases of hydatidosis reacted with 12 polypeptides of 8-116 Kda in Western immunoblot with hydatid antigens. Polypeptides of 16,24, 38,45 and 58 Kda were recognised by all hydatidosis sera but also by many sera from patients with other infections. However, polypeptides of 8 and 116 Kda were recognised by all hydatidosis sera but not by any sera from patients with cysticercosis, other parasitic infections or viral hepatitis, or from healthy controls. Thus, recognition of 8-and 116-Kda hydatid antigens by a patient's serum appears to be a specific test confirming a clinical diagnosis in an individual case of hydat idosis.

Comparative evaluation of Western Blotting in hepatic and pulmonary cystic echinococcosis

“…Our ELISA results are similar to those obtained by Zarzosa et al (1999). In our WB, we could not get sensitivity or specificity results higher than 90 % for single bands, despite the high sensitivity and specificity results obtained with antigen 5 or antigen B by others (Kanwar et al, 1992;Sbihi et al, 1997). The discrepancy may be attributed to the differences in using purified antigen or electrophoresis conditions.…”

“…Although the most sensitive band in the WB of hepatic CE, according to the ROC curve analysis, is 52-54 kDa, it seems to have little importance, in differentiating pulmonary and hepatic CE. Similarly, despite the relatively high sensitivity of the other bands of intermediate molecular weight (24 kDa, 44-46 kDa) as has been observed by others (Kanwar et al, 1992;Doiz et al, 2001), these bands did not allow differentiation between hepatic and pulmonary CE. Although some authors showed relatively high sensitivity for the 10 to 12 kDa protein, the smallest subunit of the B antigen (Verastegui et al, 1992;Doiz et al, 2001), in our investigation, this band presented lower sensitivity for detection of CE cases.…”

Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus

“…Some of the products identified in HCF by the pool of human sera in the present study may correspond to those previously described by other authors using the same antigenic extract. This may be the case of the protein of 34 kDa (recently identified by Poretti et al 1999), and the protein of 110 kDa, probably related with the component of 100 kDa described by Shambesh et al (1995), the protein of 110-120 kDa (Shapiro et al 1992), and the protein of 116 kDa (Kanwar et al 1992).…”

Dot-Elisa for evaluation of hydatid cyst wall, protoscoleces and hydatid cyst fluid antigens in the serodiagnosis of cystic echinococcosis

“…The HCF antigen is prepared as per the method described by KANWAR et al 7 . The HCF is aspirated aseptically from the fertile hydatid cyst and is collected in sterile centrifuge tube.…”