Optimization of PCR-ELISA in Detection of Human Cytomegalovirus Infection
Authors
Abstract:
Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative
similar resources
PCR optimization: Improving of Human Cytomegalovirus (HCMV) PCR to Achieve a Highly Sensitive Detection Method
Polymerase chain reaction (PCR) is a rapid and simple technique with high sensitivity and specificity. Inthe recent years, PCR has been used for rapid detection of viral nucleic acids, such as Humancytomegalovirus (HCMV), whereas, PCR optimization is an important task to be done, especially beforeit’s diagnostic application. Annealing temperature, ion concentration (especially...
full textDevelopment of a Sensitive Quantitative Competitive PCR Assay for Detection of Human Cytomegalovirus DNA
Accurate and rapid diagnosis of human cytomegalovirus (HCMV) disease in immunocompromised patients has remained as a challenge. Quantitative competitive PCR (QC-PCR) methods for detection of HCMV in these individuals have improved the positive and negative predictive values of PCR for diagnosis of HCMV disease. In this study we used QC-PCR assay, using a co-amplified DNA standard, to quantitate...
full textDetection of Leishmania major using PCR-ELISA
Background and objectives: Cutaneous leishmaniasis is endemic in most areas of Iran, and the diagnosis of its species is essential for controlling the disease. Leishmania major is the causative agent for cutaneous leishmaniasis in humans. Molecular methods are generally more sensitive than microscopic methods. The present study aimed to use a polymerase chain reaction-enzyme linked immunosorben...
full textPrevalence of Human Cytomegalovirus Infection in Iranian Prostate Cancer Patients
Background and Aim: Human cytomegalovirus (HCMV) has the potential for oncogenicity and has been associated with many malignancies, including prostate cancer. This study aimed to investigate the genomic prevalence of HCMV in prostate cancer tumors. Materials and Methods: In this case-control study, 31 patients with prostate cancer were enrolled along with 31 patients with benign prostate cance...
full textComparison of Dot-ELISA and Sandwich ELISA Diagnostic Tests in Detection of Human Hydatidosis
Hydatidosis, caused by the larval stage of Echinococcus granulosus, is one of the most important zoonosis with worldwide distribution. As its diagnosis by clinical symptoms and scanning alone is difficult and confusing, we designed the present study to achieve a sensitive and simple diagnostic method for epidemiological studies. Sera (250 samples) were collected from 90 cases of hydatidosis pro...
full textMolecular Detection of Human Cytomegalovirus in Semen of infertile Men in Tehran
Background and Aims: Human cytomegalovirus (HCMV) can induce latency and evade the immune system. The latent virus can reactive later in older ages and due to immunosuppressive conditions. Sexually transmitted diseases and viruses can influence the male reproductive system, and members of the Herpesviridae family are one of these important viruses. Regarding the importance of herpesviruses, spe...
full textMy Resources
Journal title
volume 8 issue 5
pages 14- 21
publication date 2015-01
By following a journal you will be notified via email when a new issue of this journal is published.
No Keywords
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023