A Schistosoma haematobium-Specific Real-Time PCR for Diagnosis of Urogenital Schistosomiasis in Serum Samples of International Travelers and Migrants

Abstract The study was carried out in seven reproductive herds of pigs. In three of them reproductive disorders were observed. Three herds consisted of 10-50 and four consisted of 120-500 adult sows and they were called small and medium, respectively. Fifty-seven adult sows were randomly selected from herds. Serum samples were tested using the complement fixation test and swabs from both eyes and from the vaginal vestibule were examined using real-time PCR. All serum samples were negative. Infected sows were present in each of the study herds. In total, there were 28 positive samples (53%, 28/48) in real-time PCR in sows with reproductive disorders and 35 (53%, 35/66) in sows selected from herds without problems in reproduction. One isolate proved to be Chlamydophila pecorum, whereas all the remaining were Chamydia suis

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Usefulness of a quantitative real-time PCR assay using serum samples to discriminate between inactive, serologically positive and active human brucellosis

Clinical Microbiology and Infection ◽

10.1111/j.1469-0691.2008.02095.x ◽

2008 ◽

Vol 14 (12) ◽

pp. 1128-1134 ◽

Cited By ~ 29

Author(s):

M.I. Queipo-Ortuño ◽

J.D. Colmenero ◽

M.J. Bravo ◽

M.A. García-Ordoñez ◽

P. Morata

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Human Brucellosis ◽

Quantitative Real Time Pcr ◽

Pcr Assay ◽

Serum Samples

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Rapid diagnosis of human brucellosis by SYBR Green I-based real-time PCR assay and melting curve analysis in serum samples

Clinical Microbiology and Infection ◽

10.1111/j.1469-0691.2005.01202.x ◽

2005 ◽

Vol 11 (9) ◽

pp. 713-718 ◽

Cited By ~ 40

Author(s):

M.I. Queipo-Ortuño ◽

J.D. Colmenero ◽

J.M. Reguera ◽

M.A. García-Ordoñez ◽

M.E. Pachón ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Melting Curve ◽

Rapid Diagnosis ◽

Curve Analysis ◽

Sybr Green I ◽

Melting Curve Analysis ◽

Human Brucellosis ◽

Pcr Assay ◽

Serum Samples

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Multiplex Qpcr Facilitates Identification Of Betaherpesviruses In Patients With Acute Liver Failure Of Unknown Etiology

10.21203/rs.2.9193/v1 ◽

2019 ◽

Author(s):

Jéssica Vasques Raposo ◽

Arthur Daniel Rocha Alves ◽

Alexandre Dos Santos Da Silva ◽

Damião Carlos Dos Santos ◽

Juliana Gil Melgaço ◽

...

Keyword(s):

Liver Failure ◽

Acute Liver Failure ◽

Real Time ◽

Liver Enzyme ◽

Real Time Pcr ◽

Dual Infection ◽

Unknown Etiology ◽

Hepatic Enzyme ◽

Serum Samples ◽

Multiplex Qpcr

Abstract Background: The etiology of acute liver failure (ALF) is often unknown and reported to be associated with herpesviruses in a number of cases. In this study, we examined for betaherpesviruses infections in patients with ALF of unknown etiology using a multiplex qPCR to Betaherpesviruses subfamily. Methods: Liver and serum samples from 27 patients with ALF of unknown etiology were analyzed with the aid of multiplex qPCR to identify betaherpesviruses. All positive samples were sequenced to confirm herpes infection and liver enzyme levels evaluated. Results: Betaherpesviruses infection was effectively detected using multiplex qPCR. Seven (26%), two (7%) and three (11%) cases were positive for HHV-6, HHV-7 and HCMV, respectively. Two cases of dual infection (HHV-7/HHV-6 and HHV-7/HCMV) were additionally identified. Interestingly, HHV-7 was only detected in the presence of other betaherpesviruses. Sequencing information confirmed betaherpesviruses infection. High hepatic enzyme levels and INR values>1.4 were determined in all betaherpesvirus-positive patients. Conclusions: Multiplex qPCR facilitated efficient quantification, indicating that differentiation between betaherpesviruses is possible with the sole use of real-time PCR. Liver and serum samples were positive for some betaherpesviruses, suggesting an association with ALF. Coinfection of HHV-7 with HHV-6 or HCMV was additionally detected, suggesting that the precursor betaherpesviruses infection can trigger HHV-7 infection. Based on these results, we propose that ALF patients should be screened for the presence of betaherpesviruses.

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Molecular Diagnosis of Invasive Aspergillosis and Detection of Azole Resistance by a Newly Commercialized PCR Kit

Journal of Clinical Microbiology ◽

10.1128/jcm.01032-17 ◽

2017 ◽

Vol 55 (11) ◽

pp. 3210-3218 ◽

Cited By ~ 33

Author(s):

Eric Dannaoui ◽

Frédéric Gabriel ◽

Manuel Gaboyard ◽

Gaëlle Lagardere ◽

Lucile Audebert ◽

...

Keyword(s):

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Clinical Samples ◽

Azole Resistance ◽

Rrna Gene ◽

28S Rrna ◽

Serum Samples ◽

Content Type ◽

28S Rrna Gene

ABSTRACTAspergillus fumigatusis the main species responsible for aspergillosis in humans. The diagnosis of aspergillosis remains difficult, and the rapid emergence of azole resistance inA. fumigatusis worrisome. The aim of this study was to validate the new MycoGENIEA. fumigatusreal-time PCR kit and to evaluate its performance on clinical samples for the detection ofA. fumigatusand its azole resistance. This multiplex assay detects DNA from theA. fumigatusspecies complex by targeting the multicopy 28S rRNA gene and specific TR34and L98H mutations in the single-copy-numbercyp51Agene ofA. fumigatus. The specificity ofcyp51Amutation detection was assessed by testing DNA samples from 25 wild-type or mutated clinicalA. fumigatusisolates. Clinical validation was performed on 88 respiratory samples obtained from 62 patients and on 69 serum samples obtained from 16 patients with proven or probable aspergillosis and 13 patients without aspergillosis. The limit of detection was <1 copy for theAspergillus28S rRNA gene and 6 copies for thecyp51Agene harboring the TR34and L98H alterations. No cross-reactivity was detected with various fungi and bacteria. All isolates harboring the TR34and L98H mutations were accurately detected by quantitative PCR (qPCR) analysis. With respiratory samples, qPCR results showed a sensitivity and specificity of 92.9% and 90.1%, respectively, while with serum samples, the sensitivity and specificity were 100% and 84.6%, respectively. Our study demonstrated that this new real-time PCR kit enables sensitive and rapid detection ofA. fumigatusDNA and azole resistance due to TR34and L98H mutations in clinical samples.

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Rapid quantification of hepatitis B virus DNA by direct real-time PCR from serum without DNA extraction

Journal of Medical Microbiology ◽

10.1099/jmm.0.47154-0 ◽

2007 ◽

Vol 56 (6) ◽

pp. 766-771 ◽

Cited By ~ 9

Author(s):

Zheng-Jiang Cheng ◽

Li-Hua Hu ◽

Wen-Rong Fu ◽

Yi-Rong Li

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Cost Savings ◽

Serum Samples ◽

Hepatitis B Virus Dna ◽

B Virus ◽

High Level

The purpose of this study was to quantify hepatitis B virus DNA by direct real-time PCR from serum without the need for DNA extraction. Crossing point (Cp) values were determined automatically using the second derivative maximum mode. Since serum samples from patients are inevitably haemolysed, lipaemic or icteric, the interference of endogenous substances from the serum in real-time PCR was evaluated. The result showed that, although serum protein quenched the intensity of fluorescence, the Cp value adopted to calculate the quantity of DNA copies remained unchanged. Importantly, real-time PCR from serum with or without DNA extraction reached a high level of concordance. This direct serum PCR method without the DNA extraction and gel electrophoresis allows for substantial labour and cost savings. In addition, it is also suitable for rapid DNA quantification during clinical diagnosis.

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Usefulness of real-time PCR assay targeting lipL32 gene for diagnosis of human leptospirosis in Uruguay

The Journal of Infection in Developing Countries ◽

10.3855/jidc.4110 ◽

2013 ◽

Vol 7 (12) ◽

pp. 941-945 ◽

Cited By ~ 10

Author(s):

Sabina González ◽

Juan Pablo Geymonat ◽

Elba Hernández ◽

Juan Martín Marqués ◽

Felipe Schelotto ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Dna Amplification ◽

Diagnostic Sensitivity ◽

Analytical Sensitivity ◽

Serum Samples ◽

Initial Management ◽

Negative Results ◽

Acute Infections ◽

Positive Results

Introduction: Assays based on DNA amplification can provide information that contributes to the initial management of patients with leptospirosis. However, these have not been adopted in Uruguay. Our aim was to evaluate the performance of the lipL32 real-time PCR (qPCR) for diagnosis of leptospirosis. Methodology: We analyzed by microscopic agglutination test (MAT) and lipL32 qPCR serum samples from 183 patients with suspected leptospirosis. To establish the analytical sensitivity of the qPCR, experimentally spiked samples with known amounts of Leptospira interrogans were analyzed. Results: The analytical sensitivity of the qPCR was 102 leptospires/mL. In 98 patients MAT results were negative meanwhile 85 showed positive reactions, revealing acute infections. Twenty six acute-phase sera of these 85 patients showed a positive signal by qPCR (diagnostic sensitivity 30%). In these patients the average time between onset of symptoms and collection of the first sample was 8 days. In patients with negative results for qPCR and positive MAT results (n=59) the average interval between onset of symptoms and collection of the first sample was 13 days. The qPCR did not yield false positive results. Conclusions: The qPCR had a lower diagnostic sensitivity than MAT and a higher cost. However, it allowed to make an early diagnosis in 26 patients. In patients with confirmed acute infections and negative results by qPCR, more than 8 days had elapsed between the onset of the illness and extraction of the first serum sample. Our data support that the qPCR from sera have clinical utility within the first week of illness.

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Serum Biochemistry of Lumpy Skin Disease Virus-Infected Cattle

BioMed Research International ◽

10.1155/2016/6257984 ◽

2016 ◽

Vol 2016 ◽

pp. 1-6 ◽

Cited By ~ 7

Author(s):

Murat Şevik ◽

Oğuzhan Avci ◽

Müge Doğan ◽

Ömer Barış İnce

Keyword(s):

Real Time ◽

Disease Virus ◽

Real Time Pcr ◽

Skin Disease ◽

Infected Cattle ◽

Serum Samples ◽

Lumpy Skin Disease ◽

Blood Samples ◽

Lumpy Skin Disease Virus ◽

Serum Biochemical

Lumpy skin disease is an economically important poxvirus disease of cattle. Vaccination is the main method of control but sporadic outbreaks have been reported in Turkey. This study was carried out to determine the changes in serum biochemical values of cattle naturally infected with lumpy skin disease virus (LSDV). For this study, blood samples in EDTA, serum samples, and nodular skin lesions were obtained from clinically infected animals (n=15) whereas blood samples in EDTA and serum samples were collected from healthy animals (n=15). A quantitative real-time PCR method was used to detectCapripoxvirus(CaPV) DNA in clinical samples. A real-time PCR high-resolution melt assay was performed to genotype CaPVs. Serum cardiac, hepatic, and renal damage markers and lipid metabolism products were measured by autoanalyzer. LSDV nucleic acid was detected in all samples which were obtained from clinically infected cattle. The results of serum biochemical analysis showed that aspartate aminotransferase, alkaline phosphatase, total protein, and creatinine concentrations were markedly increased in serum from infected animals. However, there were no significant differences in the other biochemical parameters evaluated. The results of the current study suggest that liver and kidney failures occur during LSDV infection. These findings may help in developing effective treatment strategies in LSDV infection.

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