scholarly journals Diagnostic Use of PCR in Carbapenamase-producing Enterobacteriaceae (CPE): An Improved Method to Overcome the presence of inhibitors for DNA Extraction from Blood Cultures

2018 ◽  
Author(s):  
Zeti Norfidiyati Salmuna ◽  
Murnihayati Hassan
Keyword(s):  
Nucleic Acid ◽  
Dna Extraction ◽  
Internal Control ◽  
Previous Method ◽  
Nucleic Acid Amplification ◽  
Culture Bottle ◽  
Predominant Genotype ◽  
Specificity And Sensitivity ◽  
Nucleic Acid Amplification Technology ◽  
Amplification Technique

AbstractBackgroundCarpanenamase-producingEnterobacteriaceae(CPE) has emerged as a threat to hospitalized patients. Phenotypic test such as Modified hodge test was less sensitive and specific especially to detectblaNDM-1 which is the most predominant genotype in this region. Nucleic acid amplification technology offers improved specificity and sensitivity. However, there is limitation in this method which is failed amplification due to the presence of inhibitors. In this study, we try to use previous method described by Villumseen et al with some modification using another DNA extraction kit.Methodology/Principle findingsThree confirmed isolates ofblaNDM-1 carbapenamase-producingKlebsiella pneumoniaewas spiked with 10 mls sterile whole blood in an aerobic Bactec Plus until the growth was detected. The blood specimen was taken and was subjected to DNA extraction method using two commercialized kits followed with multiplex PCR. All the three isolates revealedblaNDM-1 genotypes. Internal control was amplified in all three isolates.Conclusion/significanceThis study proved that we can get a specific and early diagnosis of CPE by using nucleic acid amplification technique directly from blood culture bottle and eliminate the effect of inhibitors.

scholarly journals Risk Minimization Measures for Blood Screening HIV-1 Nucleic Acid Amplification Technique Assays in Germany

2014 ◽  
Vol 41 (1) ◽  
pp. 8-8 ◽  
Author(s):  
Michael Chudy ◽  
Julia Kress ◽  
Jochen Halbauer ◽  
Margarethe Heiden ◽  
Markus B. Funk ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Amplification ◽  
Risk Minimization ◽  
Amplification Technique ◽  
Hiv 1 ◽  
Nucleic Acid Amplification Technique ◽  
Blood Screening

Role of Covid Antigen in the Diagnosis of Covid Positive in Obstetric Patients

2021 ◽  
Vol 15 (10) ◽  
pp. 3356-3358
Author(s):  
Ambreen Fatima ◽  
Nidda Yaseen ◽  
Amna Fareed ◽  
Kashif Ali Samin ◽  
Shumaela Kanwal ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Early Detection ◽  
Negative Predictive Value ◽  
Predictive Value ◽  
False Negative ◽  
Nucleic Acid Amplification ◽  
Antigen Test ◽  
Rt Pcr ◽  
Cross Sectional ◽  
Specificity And Sensitivity

Background and Aim: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapid emergence postured significant challenges on the health system in recent years. The early detection of cases is thought to be critical in preventing this pandemic by coronavirus disease (COVID-19), especially important in the obstetrical population due to theirs numerous interactions with another parturient when hospitalized for delivery. Therefore, the present study aimed to assess the COVID antigen test performance in COVID-positive obstetrics patients. Materials and Methods: This cross-sectional study was conducted on 1296 Covid-19 asymptomatic women admitted to the Obstetrics and Gynaecology Department of Muhammad Teaching Hospital & Medical College, Peshawar and Fauji Foundation Hospital, Rawalpindi for the duration of six months from February 2021 to July 2021. Antigen-based test rapid diagnostic test (RDT) was used for screening out COVID-19 positive obstetrics patients or women through nasopharyngeal swabs. Women with negative rapid antigen test results were confirmed with RT-polymers chain reaction test of nucleic acid amplification tests (NAAT). Ethical approval and informed consent were taken from the hospital ethical committee and each individual respectively. All the known positive COVID-19 patients during admission were excluded. SPSS version 24 was used for data analysis. Results: The overall prevalence of rapid antigen-positive tested patients was 13.2% (171/1296). The prevalence of positive tested women through rapid antigen test, Nucleic Acid Amplification Test (NAAT), and RT-PCR were 27 (2.1%), 51 (3.9%), and 93 (7.2%) respectively. Of the total 1296 rapid antigen tests, 27 were positive, and the false-negative confirmed positive by NAAT was 144.Thus the sensitivity of the rapid antigen test was 15.8% and the negative predictive value was 93.7%. Of the total 298 Nucleic Acid Amplification Tested had sensitivity and negative predictive value of 89.6% and 99.06% respectively. RT-PCR was carried out on 972 patients, positive diagnosed cases were 36 while 15 were initially negative and were positive with the test was repeated. The sensitivity and negative predictive value was 71.45% and 95.8% respectively. Conclusion: Our study found that Ag-RDT plays a significant role in SARS-CoV-2 early detection in infected individuals, with high specificity and sensitivity to disease infectious stage, whether symptomatic or asymptomatic, and can be used as a decision supported tool. Early detection of COVID-19 status in women admitted for delivery could benefit neonatal protection care. Keywords: Covid-19; Rapid antigen test; RT-PCR test

Quantitative nucleic acid amplification by digital PCR for clinical viral diagnostics

2016 ◽  
Vol 0 (0) ◽  
Author(s):  
Kuo Zhang ◽  
Guigao Lin ◽  
Jinming Li
Keyword(s):  
Nucleic Acid ◽  
Digital Pcr ◽  
Nucleic Acid Amplification ◽  
Target Sequence ◽  
The Past ◽  
Amplification Technique ◽  
Diagnostic Applications ◽  
Further Development ◽  
Nucleic Acid Amplification Technique ◽  
Viral Diagnostics

AbstractIn the past few years, interest in the development of digital PCR (dPCR) as a direct nucleic acid amplification technique for clinical viral diagnostics has grown. The main advantages of dPCR over qPCR include: quantification of nucleic acid concentrations without a calibration curve, comparable sensitivity, superior quantitative precision, greater resistance to perturbations by inhibitors, and increased robustness to the variability of the target sequence. In this review, we address the application of dPCR to viral nucleic acid quantification in clinical applications and for nucleic acid quantification standardization. Further development is required to overcome the current limitations of dPCR in order to realize its widespread use for viral load measurements in clinical diagnostic applications.

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