Abstract
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Improvement and Evaluation of a Staining Method for Measuring Sperm Lactate Dehydrogenase C4 Activity
by Zhaolei Cui, Cuihua Fan, Chunxiu Pang, Dezhu Zheng, Yaohua Liu, Liangyuan Chen, Duo Zhang, Fenghua Lan
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Background: This study sought to improve and evaluate a 2-hydroxyvaleric acid based staining method for detection of lactate dehydrogenase C4 (LDH-C4) activity in human spermatozoa.
Methods: A staining method for measuring sperm LDH-C4 activity with the substrate 2-hydroxyvaleric acid was improved. Expression level of LDH-C4 was assessed by Western blotting. The diagnostic performance was evaluated by plotting the receiver operating characteristic (ROC) curve.
Results: The positive products were black purple lumps concentrated in the neck segment of spermatozoa. Expression level of LDH-C4 was significantly reduced in the low activity infertile cases as compared to the matched contrasts. Decreased LDH-C4 level was significantly correlated with the declined enzyme activity (r = 0.702, p = 0.000). The ROC curve allowed for the discrimination between low and normal LDH-C4 activity cases with a sensitivity of 0.912 and specificity of 0.895, corresponding to an area under curve (AUC) of 0.941.
Conclusions: The improved method hallmarks a promising accuracy in evaluating sperm LDH-C4 activity. Downregulated LDH-C4 level is a culprit for the decreased LDH-C4 activity in spermatozoa.
DOI: 10.7754/Clin.Lab.2015.150739
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