Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis

Doiz, Olga; Benito, Rafael; Sbihi, Younes; Osuna, Antonio; Clavel, Antonio; Gómez-Lus, R

doi:10.1016/s0732-8893(01)00293-0

Cited by 48 publications

(30 citation statements)

References 24 publications

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“…The protein samples were extracted by lysis buffer (Beyotime Biotechnology, Shanghai, China) supplemented with protease inhibitors (Roche, Guangzhou, China), and the BCA TM Protein Assay Kit (Pierce, Appleton, WI, USA) was used for protein quantification. Then equal amounts of samples were separated by 10% sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred onto PVDF membranes (Millipore, Billerica, MA, USA) as described previously [24]. Specific antibodies against HIF-1a (ab179483); Caspase-9 (ab2014); cleaved Caspase-9 (ab2325); Caspase-3 (ab13847); cleaved Caspase-3 (ab49822); poly ADP ribose polymerase (PARP, ab32138); cleaved-PARP (ab32064); B-cell lymphoma-2 (Bcl-2, ab32124); serum and glucocorticoid-regulated kinase 1 (SGK1, ab59337); PI3K (ab189403); phosphorylated PI3K (p-PI3K, ab182651); AKT (ab8805); phosphorylated AKT (p-AKT, p308, ab38449; p473, ab81283); glyceraldehyde-3-posphate dehydrogenase (GAPDH) (ab8245) (all 1:1,000, Abcam, USA) were used along with appropriate fluorescent secondary antibodies (Santa Cruz biotech).…”

Section: Western Blot Assaymentioning

confidence: 99%

Inducible miR-145 expression by HIF-1α protects cardiomyocytes against apoptosis via regulating SGK1 in simulated myocardial infarction hypoxic microenvironment

et al. 2013

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Section: Western Blot Assaymentioning

confidence: 99%

Inducible miR-145 expression by HIF-1α protects cardiomyocytes against apoptosis via regulating SGK1 in simulated myocardial infarction hypoxic microenvironment

et al. 2013

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“…Alternatively, purified fractions of antigen 5 or B have been similarly tested (1,9,10,27,34,36), However, all of these antigens still exhibit some problems, mainly related to temporally delayed reactions to clinical changes (2). Thus, the persistence of specific antibodies against cyst fluid antigens for several years after recovery makes it difficult to predict the outcome of surgery early enough (5,21,29).…”

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confidence: 99%

Assessment of Echinococcus granulosus Somatic Protoscolex Antigens for Serological Follow-Up of Young Patients Surgically Treated for Cystic Echinococcosis

et al. 2008

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Echinococcus granulosus protoscolex soluble somatic antigens (PSSAs) were assessed for their prognostic value in the serological follow-up of young patients treated for cystic echinococcosis (CE), compared to conventional hydatid fluid (HF) antigen. Based on different clinical courses and outcome of infection, as well as imaging findings, patients were retrospectively classified into two different groups including either cured CE (CCE; i.e., absence of active cysts or presence of inactive cysts, respectively) and noncured CE (NCCE) patients still presenting active cysts at the end of an up to 5-year follow-up period. An immunoglobulin G (IgG)-PSSA enzyme-linked immunosorbent assay (ELISA) showed a gradual decrease in antibody levels in CCE cases, reaching seronegativity in 20% of the cases at least within 5 years postsurgery. In comparison, the conventional IgG-HF ELISA showed a significantly lower progressive decrease in antibody levels, serology becoming negative in only 15% of CCE patients at the endpoint of the follow-up period. Serological analysis of PSSA by immunoblotting yielded an interesting immunoreactive double band of 27 and 28 kDa that, in 15 (75%) of 20 CCE cases, exhibited a rapid decrease and subsequent disappearance of respective antibody reactivities within 3 years postsurgery. Conversely, anti-27-and -28-kDa antibody reactivity strongly persisted until the endpoint of the follow-up period in all of the five NCCE patients. Further analysis of the 27-and 28-kDa doublet by using affinity-purified antibodies showed that the double band was not detectable in HF. Furthermore, a predominantly IgG4 subclass-restricted humoral immune response against the 27-and 28-kDa antigens was demonstrated in seroreactive CE patients. Overall, an anti-27-and -28-kDa response appeared to correlate with cyst activity. In conclusion, PSSA represents a useful candidate to carry out a serologic follow-up of CE subsequent to treatment and deserves further respective evaluation for other age groups of CE patients.

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“…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.…”

Section: Discussionmentioning

confidence: 67%

“…But, their diagnostic value in suspected cases of CE is limited because of the variable sensitivity and specificity (Carmena et al, 2006). Therefore, use of confirmation test such as WB (Western Blotting) is very important in serodiagnosis of CE (Doiz et al, 2001). The hydatid cysts of E. granulosus tend to form in the liver (50-70 %) or lung (20-30 %) but may be found in any organ of the body (< 10 %), including brain, heart and bones (Schantz & Gottstein, 1986).…”

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Comparative evaluation of Western Blotting in hepatic and pulmonary cystic echinococcosis

Akısü

Delıbaş

Biçmen³

et al. 2006

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Summary :Many serological tests are widely used in the diagnosis of cystic echinococcosis (CE), caused by the larval stages of Echinococcus granulosus. The present study was carried for differentiation between hepatic and pulmonary cystic echinococcosis by Western Blotting (WB). A total of 121 sera from patients with hepatic CE (37), pulmonary CE (31) and controls (53; consisting of six healthy, seven Hymenolepis nana infection, 20 hepatic and 20 pulmonary diseases other than CE) were examined. In all of the CE patients, E. granulosus infection was confirmed by surgical intervention. Sera were previously tested using IHA and ELISA to detect the E. granulosus specific antibodies. Sera from hepatic cases of CE reacted with 16 polypeptides of 6-116 kDa and sera from pulmonary cases of CE reacted with 14 polypeptides of 4-130 kDa by Western Blotting. The WB test enabled the detection of antibodies in the hepatic CE samples for proteins of 24, 32-34, 44-46 and 52-54 kDa in molecular weight in 78.4 %, 75.7 %, 78.4 % and 89.2 % of the patients, respectively. In the pulmonary CE samples sera WB test enabled the detection of antibodies 24, 44-46, 100, 110, 116 and 120-124 kDa in molecular weight in 81.3 %, 75.0 %, 87.5 %, 71.9 %, 84.4 % and 65.6 % of the patients, respectively. We indicated that the antigenic components of high molecular weight can be good candidates for differentiation of hepatic CE from pulmonary CE. de 24,[32][33][34][44][45][46]4 %, 75,7 %, 78,4 % et 89,2 % des patients, respectivement, et, en cas de KH pulmonaire, de protéines de 24,[44][45][46] 100, 110,[116][117][118][119][120][121][122][123][124]3 %, 75,0 %, 87,5 %, 71,9 %, 84,4 % et 65,6 Résumé

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Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis

Cited by 48 publications

References 24 publications

Inducible miR-145 expression by HIF-1α protects cardiomyocytes against apoptosis via regulating SGK1 in simulated myocardial infarction hypoxic microenvironment

Inducible miR-145 expression by HIF-1α protects cardiomyocytes against apoptosis via regulating SGK1 in simulated myocardial infarction hypoxic microenvironment

Assessment of Echinococcus granulosus Somatic Protoscolex Antigens for Serological Follow-Up of Young Patients Surgically Treated for Cystic Echinococcosis

Comparative evaluation of Western Blotting in hepatic and pulmonary cystic echinococcosis

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