scholarly journals Rapid identification of melioidosis agent by an insulated isothermal PCR on a field–deployable device

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9238
Author(s):  
Kek Heng Chua ◽  
E. Wei Tan ◽  
Hwa Chia Chai ◽  
SD Puthucheary ◽  
Ping Chin Lee ◽  
...  
Keyword(s):  
Burkholderia Pseudomallei ◽  
Clinical Laboratory ◽  
Rapid Identification ◽  
Chain Reaction ◽  
Serious Illness ◽  
Lower Detection Limit ◽  
Lower Detection ◽  
Polymerase Chain ◽  
Intracellular Motility ◽  
Insulated Isothermal Pcr

Background Burkholderia pseudomallei causes melioidosis, a serious illness that can be fatal if untreated or misdiagnosed. Culture from clinical specimens remains the gold standard but has low diagnostic sensitivity. Method In this study, we developed a rapid, sensitive and specific insulated isothermal Polymerase Chain Reaction (iiPCR) targeting bimA gene (Burkholderia Intracellular Motility A; BPSS1492) for the identification of B. pseudomallei. A pair of novel primers: BimA(F) and BimA(R) together with a probe were designed and 121 clinical B. pseudomallei strains obtained from numerous clinical sources and 10 ATCC non-targeted strains were tested with iiPCR and qPCR in parallel. Results All 121 B. pseudomallei isolates were positive for qPCR while 118 isolates were positive for iiPCR, demonstrating satisfactory agreement (97.71%; 95% CI [93.45–99.53%]; k = 0.87). Sensitivity of the bimA iiPCR/POCKIT assay was 97.52% with the lower detection limit of 14 ng/µL of B. pseudomallei DNA. The developed iiPCR assay did not cross-react with 10 types of non-targeted strains, indicating good specificity. Conclusion This bimA iiPCR/POCKIT assay will undoubtedly complement other methodologies used in the clinical laboratory for the rapid identification of this pathogen.

scholarly journals A comparative evaluation of two chromogenic media for surveillance of carbapenem-resistant Enterobacterales with non-carbapenemase-producing or carbapenemase-producing strains other than blaKPC: blaGES-5, blaNDM-1 or blaVIM-2

2019 ◽  
Vol 43 (3) ◽  
pp. 173-176
Author(s):  
Chang-Hun Park
Keyword(s):  
Rapid Detection ◽  
Comparative Evaluation ◽  
Susceptibility Testing ◽  
Rapid Identification ◽  
Surveillance Program ◽  
Contact Precaution ◽  
Chain Reaction ◽  
Carbapenem Resistant ◽  
Control And Prevention ◽  
Polymerase Chain

Abstract Background Infections caused by carbapenem-resistant Enterobacterales (CREs) are an emerging problem associated with high rates of morbidity and mortality. CREs are divided into two categories (carbapenemase-producing [CP] CRE and non-CP CRE). The most prevalent carbapenemase produced by Enterobacterales is Klebsiella pneumoniae carbapenemase (KPC) in Korea. Rapid identification of CREs is clinically important in infection control precaution. We compared the performance of two chromogenic media (chromID CARBA agar and CHROMagar KPC agar) for non-CP CREs or CP CREs with blaGES-5, blaNDM-1 or blaVIM-2 in a Korean hospital. Methods The study was carried out during a 3-month period from April to June 2017 during the surveillance program for CRE colonization. Antimicrobial susceptibility testing (AST) and polymerase chain reaction (PCR) were performed at the Korean Centers for Disease Control and Prevention. Results A total of 45 rectal swabs from 42 hospitalized patients were examined. Sensitivity of both chromID CARBA and CHROMagar KPC were 100% for CP CREs; and 50% and 100% for non-CP CREs, respectively. Specificity of chromID CARBA and CHROMagar KPC were 89.2% and 70.3% for CP CRE, respectively; and 76.9% and 66.7% for non-CP CRE, respectively. Conclusions The CHROMagar KPC is useful to monitor non-CP and CP CREs. The chromID CARBA is efficient for rapid detection of CP CREs requiring high contact precaution.

Rapid identification of benomyl resistant strains of Botrytis cinerea using the polymerase chain reaction

1995 ◽  
Vol 99 (12) ◽  
pp. 1483-1488 ◽  
Author(s):  
Joanne E. Luck ◽  
Michael R. Gillings
Keyword(s):  
Polymerase Chain Reaction ◽  
Botrytis Cinerea ◽  
Rapid Identification ◽  
Chain Reaction ◽  
Resistant Strains ◽  
Polymerase Chain

Rapid identification of the common apo E isoform genotype using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP)

1994 ◽  
Vol 8 (1) ◽  
pp. 51-54 ◽  
Author(s):  
Ryoji Aozaki ◽  
Ryuji Kawaguchi ◽  
Utako Ogasa ◽  
Kazumasa Hikiji ◽  
Nobuhiko Kubo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Single Strand Conformation Polymorphism ◽  
Rapid Identification ◽  
Single Strand ◽  
Chain Reaction ◽  
Apo E ◽  
The Common ◽  
Polymerase Chain ◽  
Conformation Polymorphism

scholarly journals Methods for identification of causative agents of dangerous and particularly dangerous infections based on the analysis of nucleic acids

2018 ◽  
Vol 2 (4) ◽  
pp. 22-35
Keyword(s):  
Nucleic Acids ◽  
Dna Amplification ◽  
Rapid Identification ◽  
Chain Reaction ◽  
Biological Contamination ◽  
Laboratory Facilities ◽  
Trained Personnel ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Goals And Objectives

One of the main tasks of the NBC Protection Troops is accurate and rapid identification of infectious disease causative agents in case of establishing the fact of biological contamination. Different methods based on the analysis of nucleic acids are most preferred for this purpose. Most of them are based on DNA amplification by polymerase chain reaction (PCR). The result is detected by electrophoretic separation of amplification products, as well as by registration of endpoint fluorescent signal (FLASH modification) or in real time (PCR-RT). Other methods of DNA amplification, such as ligase chain reaction (LCR) and isothermal amplification, are also applicable in practice. The article also describes some identification methods based on nucleic acid sequencing: multilocus sequence typing (MLST) method, sequencing of individual genes and complete genome sequencing. It is concluded that the choice of identification method should be based on the goals and objectives, laboratory facilities, availability of trained personnel and funding levels. Despite the fact that the most informative are methods based on sequencing nucleotide sequences, their implementation in the field is difficult so far due to technological requirements

Direct colony polymerase chain reaction for rapid identification of yeasts isolated from blood specimen

2016 ◽  
Vol 18 (2) ◽  
pp. 91 ◽  
Author(s):  
AnupmaJ Kindo ◽  
RajyoganandhS Vijayaraman ◽  
Vijayakumar Ramaraj
Keyword(s):  
Polymerase Chain Reaction ◽  
Rapid Identification ◽  
Chain Reaction ◽  
Colony Polymerase Chain Reaction ◽  
Blood Specimen ◽  
Polymerase Chain

scholarly journals Tuberculosis osteomyelitis of the tibia mimicking Brodie abscess: A case report and review of the literature

2019 ◽  
Vol 7 ◽  
pp. 2050313X1986945
Author(s):  
Abdülkadir Sari ◽  
Yaşar Mahsut Dinçel ◽  
Ibrahim Halil Erdogdu ◽  
Hakan Sezgin Sayıner ◽  
Ismail Agir ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Laboratory ◽  
Histopathological Examination ◽  
Bone Destruction ◽  
Rare Condition ◽  
Chain Reaction ◽  
Laboratory Findings ◽  
Radiographic Appearance ◽  
Brodie’S Abscess ◽  
Polymerase Chain

Background: Tuberculosis osteomyelitis is rarely seen in the diaphyseal bones. It may be confused with Brodie’s abscess due to similar clinical, radiological and laboratory findings. Late diagnosis of the disease causes bone destruction. Tuberculosis osteomyelitis of the bone is a rare condition caused by the Mycobacterium tuberculosis. Its incidence has increased in Western countries in recent years due to HIV infection, increasing elderly population and emerging resistant strains. The slow progress of tuberculous osteomyelitis, due to lack of significant elevations in the laboratory values and changes in the radiographic appearance, often leads to confusion with the subtypes of subacute osteomyelitis, defined as Brodie’s abscess. These two low-virulence clinical cases often lead to delays in diagnosis and progressive bone destruction. Case presentation: We report a 65-year-old male patient who presented to our clinic with pain, swelling and sensitivity in the left leg. Diagnosed with infection in the tibia, the patient had undergone antibiotherapy. However, the patient’s symptoms were not resolved and we performed bone curettage and cementation. M. tuberculosis-specific DNA was detected by real-time polymerase chain reaction and the M. tuberculosis complex was produced from the perioperative samples. Conclusion: In conclusion, histopathological examination and polymerase chain reaction are essential before surgery of subacute and chronic osteomyelitis with atypical clinical, laboratory and radiological findings for early diagnosis and accurate treatment.

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