Related Experiment Video
Updated: Jun 25, 2026

Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Hepatitis C virus detection in the semen of infected patients
Norma de Paula Cavalheiro1, Ana Carolina de Oliveira Santos, Carlos Eduardo Melo
1Department of Infectious Diseases, Clinical Hospital, University of São Paulo, SP, Brazil. norma@usp.br
Insights
Hepatitis C virus (HCV) RNA was investigated in patient semen. While blood tested positive, most semen samples showed negative results, suggesting potential false negatives in HCV detection within seminal fluid.
Area of Science:
- Virology
- Hepatology
- Public Health
Background:
- Hepatitis C virus (HCV) infection remains a significant global health concern.
- Understanding HCV transmission routes, particularly sexual transmission and via seminal fluid, is crucial but incompletely elucidated.
- The presence and detectability of HCV in semen require further investigation.
Purpose of the Study:
- To determine the presence of Hepatitis C virus RNA in the seminal fluid of infected patients.
- To analyze HCV RNA in different semen fractions: seminal plasma, leukocytes, and spermatozoa.
- To evaluate the reliability of detection methods for HCV in semen samples.
Main Methods:
- Thirteen patients with confirmed HCV infection were enrolled.
- Semen was fractionated into seminal plasma, leukocytes, and spermatozoa using Percoll gradients.
- HCV RNA was extracted and detected using the Roche Amplicor assay and agarose gel electrophoresis.
Main Results:
- All blood samples tested positive for HCV RNA.
- The Roche Amplicor assay detected inhibition in 87% of total semen samples and fractions.
- Only one spermatozoon fraction tested positive; all other semen samples (total, plasma, leukocytes) were negative, indicating potential false-negative results.
Conclusions:
- Standard HCV detection methods may yield false-negative results in semen samples.
- Further research is needed to optimize HCV detection in seminal fluid.
- The findings highlight challenges in assessing sexual transmission of HCV due to detection limitations.
Abstract:
Though HCV infection is a serious public health problem, some aspects of its biology are still not well understood, such as its transmission through seminal fluid and sexual transmission. We looked for HCV in the semen of infected patients. Thirteen patients were included. Semen fractions (seminal plasma, leukocytes and spermatozoa) were separated with 45% and 90% Percoll gradients. The HCV-RNA in blood and semen fractions was extracted using the same protocol (AMPLICOR Roche) and was detected using the qualitative Roche Amplicor test and by agarose gel electrophoresis, with ethidium bromide staining. The mean age of the patients was 40.7 years. Risk factors for the acquisition of HCV included injectable and inhaled drug use in six (42.8%), blood transfusion in four (28.6%), and no risk factors in four (28.6%) patients. Genotype 1 was detected in 62% of the patients, followed by genotype 3 in 23% and genotype 2 in 15%. All blood samples were positive, regardless of the technique used for detection. All semen samples identified by Roche Amplicor and analyzed by agarose gel electrophoresis were negative. Among the 52 semen samples (total and fractions) identified by the Roche Amplicor method, 45 (87%) were inhibited. A negative result was recorded for one (1.9%) total semen sample, one (1.9%) leukocyte and four (7.7%) seminal plasma fractions. Only one (1.9%) sample of the spermatozoon fraction was positive. The results obtained suggested false-negative reactions for the semen samples.
Related Concept Videos
Hepatitis
Cytomegalovirus Disease
Viral Hepatitis I: Introduction

