Improvement and Evaluation of a Staining Method for Measuring Sperm Lactate Dehydrogenase C4 Activity
| Autor: | Yaohua Liu, De-Zhu Zheng, Fenghua Lan, Zhaolei Cui, Liangyuan Chen, Chunxiu Pang, Duo Zhang, Cuihua Fan |
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| Rok vydání: | 2016 |
| Předmět: |
Adult
Male medicine.medical_specialty Improved method Biology General Biochemistry Genetics and Molecular Biology Male infertility chemistry.chemical_compound Internal medicine Lactate dehydrogenase medicine Humans Receiver operating characteristic L-Lactate Dehydrogenase Staining and Labeling Temperature Hydrogen-Ion Concentration Middle Aged medicine.disease Sperm Spermatozoa Enzyme assay Staining Blot Isoenzymes Endocrinology chemistry ROC Curve biology.protein |
| Zdroj: | Clinical laboratory. 62(4) |
| ISSN: | 1433-6510 |
| Popis: | 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. Down-regulated LDH-C4 level is a culprit for the decreased LDH-C4 activity in spermatozoa. |
| Databáze: | OpenAIRE |
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