Key result
The ISOL reaction yielded significantly fewer false-positive results for detecting myocyte apoptosis (5%) compared with ISEL (22%) or TUNEL (22%) (P=0.027).
Why the study?
Does the ISOL reaction reduce false-positive results compared to ISEL and TUNEL when detecting myocyte apoptosis in human heart tissue?
Observational (n=41)
Does the ISOL reaction reduce false-positive results compared to ISEL and TUNEL when detecting myocyte apoptosis in human heart tissue?
The ISOL reaction yields significantly fewer false-positive results than TUNEL or ISEL for detecting cardiomyocyte apoptosis in human heart tissue, suggesting it may be a more reliable method.
ISOL may reduce false positives for myocyte apoptosis in animal models; leaves open validation and utility in human cardiac tissue.
We assessed the efficiency of detecting myocyte apoptosis within human hearts using in situ enzymatic reactions in paraffin-embedded tissue samples: in situ end labeling (ISEL), terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling (TUNEL), and in situ oligoligation (ISOL). The reactions were carried out in explanted hearts (idiopathic dilatative cardiomyopathy, n = 6; ischemic heart disease, n = 3) and in endomyocardial biopsy specimens (EMBs; n = 32) obtained from transplanted human hearts. The results were verified by DNA laddering. The ISOL reaction led to a significantly (P = .027) smaller number of false-positive results (2/41 [5%]) compared with assessment by ISEL (9/41 [22%]) or TUNEL (9/41 [22%]). Only 1 ISEL+ apoptotic cardiomyocyte was found in specimens from explanted hearts. Among the EMBs, 1 specimen had TUNEL+ apoptotic cardiomyocytes and 1 specimen had ISEL+ apoptotic cardiomyocytes. This implies that verifying results by independent methods must be used for TUNEL and ISEL techniques. A smaller number of false-positive results makes interpretation of ISOL results easier, although the sensitivity of this reaction remains to be established.
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Lesauskaitė et al. (2004) conducted an observational in Heart failure and heart transplantation (n=41). In situ oligoligation (ISOL) vs. ISEL and TUNEL was evaluated on False-positive results for detecting myocyte apoptosis (p=0.027). The ISOL reaction yielded significantly fewer false-positive results for detecting myocyte apoptosis (5%) compared with ISEL (22%) or TUNEL (22%) (P=0.027).
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