scholarly journals APPLICATION OF ELISA-IGM (MAC-ELISA) FOR DETERMINING ETIOLOGICAL LINK VIRUS TYPES IN CONCRET CASES OF THE DISEASE

2017 ◽  
Vol 22 (3) ◽  
pp. 116-121
Author(s):  
Yuliya A. Akinshina ◽  
V. F Larichev ◽  
M. A Sayfullin ◽  
S. G Mardanly ◽  
A. M Butenko
Keyword(s):  
Dengue Fever ◽  
Rt Pcr ◽  
Serum Samples ◽  
Dengue Viruses ◽  
Etiological Role ◽  
Igm Elisa ◽  
Pcr Method

105 sera from 101 patients with a confirmed diagnosis of dengue fever (LD) were examined using monospecific ELISA-IgM test. Monospecific results were observed in 39 samples (37.1%). 27 sample of them were taken during the first 7 days of the disease. In those systems were examined 23 serum samples of 23 patients, in which etiological role of one of the four dengue viruses was been established by the RT-PCR method. In this case, the coincidence-IgM ELISA results and the RT-PCR took place in 14 sera (60.9%).

scholarly journals Typing of Dengue Viruses in Clinical Specimens and Mosquitoes by Single-Tube Multiplex Reverse Transcriptase PCR

1998 ◽  
Vol 36 (9) ◽  
pp. 2634-2639 ◽  
Author(s):  
Eva Harris ◽  
T. Guy Roberts ◽  
Leila Smith ◽  
John Selle ◽  
Laura D. Kramer ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Dengue Fever ◽  
Internal Control ◽  
Extraction Procedure ◽  
Rna Degradation ◽  
Viral Rna ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Single Tube ◽  
Dengue Viruses

In recent years, dengue viruses (serotypes 1 to 4) have spread throughout tropical regions worldwide. In many places, multiple dengue virus serotypes are circulating concurrently, which may increase the risk for the more severe form of the disease, dengue hemorrhagic fever. For the control and prevention of dengue fever, it is important to rapidly detect and type the virus in clinical samples and mosquitoes. Assays based on reverse transcriptase (RT) PCR (RT-PCR) amplification of dengue viral RNA can offer a rapid, sensitive, and specific approach to the typing of dengue viruses. We have reduced a two-step nested RT-PCR protocol to a single-tube reaction with sensitivity equivalent to that of the two-step protocol (1 to 50 PFU) in order to maximize simplicity and minimize the risk of sample cross-contamination. This assay was also optimized for use with a thermostable RT-polymerase. We designed a plasmid-based internal control that produces a uniquely sized product and can be used to control for both reverse transcription or amplification steps without the risk of generating false-positive results. This single-tube RT-PCR procedure was used to type dengue viruses during the 1995 and 1997-1998 outbreaks in Nicaragua. In addition, an extraction procedure that permits the sensitive detection of viral RNA in pools of up to 50 mosquitoes without PCR inhibition or RNA degradation was developed. This assay should serve as a practical tool for use in countries where dengue fever is endemic, in conjunction with classical methods for surveillance and epidemiology of dengue viruses.

A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples

2008 ◽  
Vol 153 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Nadine Sadon ◽  
Anne Delers ◽  
Richard G. Jarman ◽  
Chonticha Klungthong ◽  
Ananda Nisalak ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Sensitive Detection ◽  
Rt Pcr ◽  
Serum Samples ◽  
Pcr Method

scholarly journals Molecular and Haematological Analysis of DENV-3 Among Children in Lahore, Pakistan

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Muhammad Kashif ◽  
Muhammad Afzal ◽  
Basit Zeshan ◽  
Hasnain Javed ◽  
Salma Batool ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Rna Virus ◽  
Tertiary Care ◽  
Screening Method ◽  
Time Interval ◽  
Short Time Interval ◽  
Rt Pcr ◽  
Serum Samples ◽  
Hematological Analysis

Background: Dengue virus (DENV) is an RNA virus belonging to the family Flaviviridae of the genus Flavivirus with worldwide distribution. Dengue fever is caused by any of four closely-related serotypes DENV, an emerging pandemic-prone viral disease in many regions of the world. Objectives: The present study aimed to determine the prevalence of dengue virus genotypes and serotypes in children aged below 15 years in Lahore, Pakistan. Methods: In this study, 112 serum samples were collected from clinically suspected dengue fever patients from March 2017 to December 2018 at different tertiary care hospitals in Lahore, Pakistan. Regarding the patients’ age, the samples were divided into four groups from A to D (i.e., 0 - 1, 1 - 5, 5 - 10, and 10 - 15 years of age). Rapid immuno-chromatography (ICT) test was conducted on the collected serum samples, followed by quantitative RT-PCR for serotype of dengue virus. Results: Out of 112 samples, 34 samples were diagnosed as DENV positive by the rapid ICT screening method. No virus was detected in groups A and B, while three samples were positive in group C (1 boy and two girls), and 31 samples (23 boys and 8 girls) were positive in group D. The results of quantitative RT-PCR exclusively showed DEN-3 serotype in all the ICT positive samples. The results indicated that the prevalence of DEN-3 serotype in children was 100%, indicating that DEN-3 serotype might cause severe epidemics in the future in Lahore, Pakistan. Hematological analysis revealed an increase in hematocrits in 41.1% dengue-positive cases. Leucopenia was prominent in 79.4% of the cases, while Thrombocytopenia was reported in 70.5% of the participants. The biochemical analysis also indicated an increase in liver enzymes in patients (ALT 88%, AST 79%), while the lower levels of cholesterol (69 %) and serum albumin (25%) were also observed. Conclusions: Dengue virus spreads and grows quickly worldwide over a highly short time interval. Dengue fever claims for a significant number of lives. This study would help individuals know about the status of laboratory parameters in dengue fever and detect how to overcome the prevalence of Dengue virus.

scholarly journals Evaluation of NS1, IgM ELISA and RT-PCR in diagnosis of dengue fever

2018 ◽  
Vol 6 (7) ◽  
pp. 2440
Author(s):  
Deepak Kumar ◽  
Rajesh Kumar Verma ◽  
Amit Singh ◽  
Manoj Kumar ◽  
Dharmendra Prasad Singh ◽  
...  
Keyword(s):  
Mortality Rate ◽  
Dengue Fever ◽  
Sensitivity And Specificity ◽  
Late Phase ◽  
Accurate Diagnosis ◽  
Suspected Case ◽  
Rt Pcr ◽  
Igm Elisa ◽  
Antigen Elisa ◽  
Ns1 Antigen

Background: Dengue fever is a mosquito borne disease caused by flavivirus. Its cases are increasing in India with increasing mortality rate year by year hence, prompt and accurate diagnosis is necessary to prevent morbidity and mortality.Methods: In this study we enrolled 125 clinically suspected cases of dengue. All the collected samples were processed for RT-PCR, NS1 and IgM ELISA. We evaluated NS1 antigen ELISA alone, and combination of NS1 and IgM ELISA against RT-PCR.Results: Among 125 clinically suspected case 67 were positive by RT-PCR and 58 were negative. Sensitivity and Specificity of NS1 ELISA and NS1 with IgM ELISA (in combination) against RT-PCR were 83.58%, 94.82% and 95.55%, 79.31% respectively. (p<0.001).Conclusions: The NS1 ELISA alone was sufficient to detect acute phase of dengue fever, although, combination of NS1 and IgM proved to be most appropriate method for detection of acute as well as late phase of dengue fever.

scholarly journals All four dengue virus serotypes co-circulate in concurrent dengue infections in a single dengue session in Chittagong, Bangladesh

2021 ◽  
Vol 8 (1) ◽  
pp. 1042-1048
Author(s):  
Moushumi Ghosh Roy ◽  
Kutub Uddin ◽  
Din Islam ◽  
Anjuvan Singh ◽  
Mohammad Monirul Islam
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Disease Severity ◽  
Clinical Symptoms ◽  
Global Public Health ◽  
Rt Pcr ◽  
Serum Samples ◽  
Dengue Disease ◽  
Denv Serotypes ◽  
Public Health Burden

Purposes: Dengue fever, a mosquito-borne viral disease, is a global public health burden affecting millions of people each year and over 40% of world populations are at risk of dengue. Therefore, prompt and accurate dengue diagnosis is inevitable for disease surveillance and for aiding disease management. In this study we report dengue virus (DENV) seroprevalence in Chittagong, Bangladesh along with clinical manifestation of dengue infections. Methods: All samples included in this study were selected based on dengue NS1-based diagnosis, clinical sign and symptoms were judged by expert clinical physicians and infecting DENV serotyping was done by RT-PCR. The blood cells (Platelet, Haematocrit, WBC etc) were analyzed using Haematology cell counter. Results: First, among the 112 DENV infected serum samples tested by RT-PCR, 42 were DENV positive where 76% samples had single DENV serotype infection and 24% were concurrently infected with two or more DENV serotypes, indicating that all four DENVs were present in a single dengue session in Chittagong, Bangladesh. Then, DENV4 was the most prevailed serotype, followed by DENV2, DENV1 and DENV3 in single DENV serotype infections. However, in almost 90% cases of concurrent multiple DENV infections DENV1 serotype was present. A detail analysis of clinical data clearly indicated that DENV1 and DENV2 resulted very similar patterns of clinical symptoms which were quite different from those caused by DENV3 and DENV4. For example, ache and pain were absent in DENV3 infection and diarrhea was absent in DENV4 infections. Furthermore, DENV3, both in single and concurrent multiple DENV infections, might increase dengue disease severity as observed highly reduced platelet counts along with increased WBC in patients infected with DENV3 serotype. Conclusion: All four DENV serotypes, both as single and concurrent multiple DENV infections, were present in single dengue session in Bangladesh. Despite having very similar sequences and structures all four DENVs might produce different disease spectra, ranging from classical dengue fever to dengue hemorrhagic fever. Concurrent multiple DENV infections could contribute increased dengue disease severity in dengue outbreaks in Bangladesh. Bioresearch Commu. 8(1): 1042-1048, 2022 (January)

scholarly journals Comparison of Cytokine Expression Profile in Chikungunya and Dengue Co-Infected and Mono-Infected Patients’ Samples

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 166
Author(s):  
Saravana Murali Krishnan ◽  
Jayashri Mahalingam ◽  
Shanthi Sabarimurugan ◽  
Thiruvengadam Muthu ◽  
Baskar Venkidasamy ◽  
...  
Keyword(s):  
Viral Load ◽  
Expression Profiles ◽  
Febrile Illness ◽  
Cytokine Expression ◽  
Rt Pcr ◽  
Serum Samples ◽  
Cytokine Expression Profile ◽  
Significant Difference ◽  
Ifn Γ

Chikungunya (CHIKV) and Dengue (DENV) viruses cause an acute febrile illness which is hard to clinically differentiate and treat since both exhibit similar symptoms. Hence, this study was aimed at identifying the expression profiles of cytokines on co-infected samples and compare with CHIKV and DENV mono-infected samples. Serum samples of 292 suspected patients during 2009–2011 were analyzed. The presence of primary (IgM)/secondary (IgG) antibodies and early NS1 Dengue antigens were confirmed by capture ELISA. Molecular diagnosis and serotypes were discriminated by RT-PCR, confirmed by sequencing. All the plasma samples were assayed for cytokine expression by BDTM cytometry bead array (CBA) and compared with independent mono-infection viral load. Among the tested samples, 82 were confirmed as Dengue positive; 52 through IgM (17.8%), and 30 through IgG (10.2%). Additionally, 186 samples were confirmed as Chikungunya, 96 through IgM (32.6%) and 92 through IgG (31.5%) ELISA, respectively. Interestingly, 19 patients were co-infection positive in which, only 6 were confirmed for CHIKV and 7 for DENV by RT-PCR. Among 8 cytokines, IL-2, IL-8, IFNα, IFN γ, and IL-12 were found to be significantly different between co-infected and CHIKV mono-infected patients and correlated with viral load. DENV viral load was correlated with cytokine expression and a significant difference in IL-2 and IL-12 was observed between DENV mono-infected and DENV and CHIKV co-infected patients. Results indicated that apart from serological and molecular confirmation, cytokines could be used as a specific biomarker for the diagnosis of DENV and CHIKV. In the future, the role of independent cytokines can be determined to understand the pathogenesis and etiology of these dreadful diseases.

scholarly journals Detection of Hepatitis C Virus Coinfection in Patients with Dengue Diagnosis

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Carlos Machain-Williams ◽  
Lourdes Talavera-Aguilar ◽  
Rosa Carmina Cetina-Trejo ◽  
Jaquelin Carrillo-Navarrete ◽  
Nubia Rivero-Cárdenas ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Serological Test ◽  
Clinical Manifestations ◽  
Rt Pcr ◽  
Serum Samples ◽  
Viral Isolation ◽  
Igm Elisa ◽  
Male Patients ◽  
Hcv Coinfection

Coinfection produced by dengue virus (DENV) and hepatitis C virus (HCV) is a serious problem of public health in Mexico, as they both circulate in tropical zones and may lead to masking or complicating symptoms. In this research, we detected active coinfected patients by HCV residing in the endemic city of Mérida, Yucatán, Mexico, with positive diagnosis to dengue during the acute phase. We performed a retrospective analysis of 240 serum samples from dengue patients. The IgM-ELISA serological test was used for dengue diagnosis, as well as viral isolation to confirm infection. DENV and HCV were detected by RT-PCR. Thus, 31 (12.9%) samples showed DENV-HCV coinfection, but interestingly the highest frequency of coinfection cases was found in male patients presenting hemorrhagic dengue in 19/31 (61.29%), with a predominance of 12 : 7 in males. Firstly, coinfection of DENV-HCV in Mérida, Mexico, was detected in young dengue patients, between 11 and 20 years old (38.7%), followed by those between 21 and 30 years old (32%); only 16.13% were between 0 and 10 years of age. Diagnosis of HCV infection in patients with dengue is highly recommended in order to establish potential risk in clinical manifestations as well as dictate patients' special care.

TO DETERMINE THE ROLE OF CA125 IN THE DIAGNOSIS OF ACUTE APPENDICITIS

2020 ◽  
Vol 4 (7) ◽  
Author(s):  
Vinod Kumar ◽  
Bhupen Songra ◽  
Richa Jain ◽  
Deeksha Mehta
Keyword(s):  
Acute Appendicitis ◽  
Clinical Diagnosis ◽  
Tertiary Care Hospital ◽  
Tertiary Care ◽  
Ca 125 ◽  
Care Hospital ◽  
Serum Samples ◽  
Roc Curve Analysis ◽  
Surgery Department

Background: the present study was under taken to determine the role of CA-125 in the diagnosis of acute appendicitis (AA), to prevent its complications and also in preventing negative appendicectomies in tertiary care hospital. Methods: The study was conducted at a tertiary care and research center between 01/03/2018 to 30/06/2019. Patients admitted to the surgery department with diagnosis of AA were considered for the study. After informed consent, a, standardized history was obtained as a case Performa. Serum samples from all the cases with clinical diagnosis of AA were obtained and stored. Only the cases with histopathologically approved AA were included in the study. Cases operated for clinical diagnosis of AA, but not histopathologically proven AA was not included in the study. CA125 levels in cases with definitive diagnosis of AA were measured. Results: In present study, ROC curve analysis revealed the sensitivity of 87.27 % and specificity of 90.91 % when the CA 125 cut-off value of > 16.8 was taken to diagnose acute appendicitis. AUC was 0.911 with a standard error of 0.0292. Conclusion: In this study we have observed that CA125 showed a positive correlation with acute appendicitis, that was statistically not significant (P>0.05). We didn’t evaluate the correlation with the disease severity. We consider that CA125 can be used as a marker in acute appendicitis cases although further research is still needed. Keywords: CA125, Acute Appendicitis, Surgery.

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