Simultaneous species identification and detection of methicillin resistance in staphylococci using triplex real-time PCR assay

2006 ◽  
Vol 56 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Negar Shafiei Sabet ◽  
Geetha Subramaniam ◽  
Parasakthi Navaratnam ◽  
Shamala Devi Sekaran
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Methicillin Resistance ◽  
Pcr Assay

Species Identification of Fox-, Mink-, Dog-, and Rabbit-Derived Ingredients by Multiplex PCR and Real-Time PCR Assay

2017 ◽  
Vol 185 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Qingqing Wu ◽  
Shengnan Xiang ◽  
Wenjun Wang ◽  
Jinyan Zhao ◽  
Jinhua Xia ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Pcr Assay

scholarly journals Detection and species identification of microsporidial infections using SYBR Green real-time PCR

2011 ◽  
Vol 60 (4) ◽  
pp. 459-466 ◽  
Author(s):  
Spencer D. Polley ◽  
Samuel Boadi ◽  
Julie Watson ◽  
Alan Curry ◽  
Peter L. Chiodini
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Enterocytozoon Bieneusi ◽  
Sybr Green ◽  
Pcr Assay ◽  
Real Time Analysis ◽  
Highly Sensitive ◽  
Stool Samples

Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron microscopy. This study describes a single SYBR Green real-time PCR assay for the simultaneous detection and species identification of such infections. This assay was highly sensitive, routinely detecting infections containing 400 parasites (g stool sample)−1, whilst species identification was achieved by differential melt curves on a Corbett Life Science Rotor-Gene 3000. A modification of the QIAamp DNA tissue extraction protocol allowed the semi-automated extraction of DNA from stools for the routine diagnosis of microsporidial infection by real-time PCR. Of 168 stool samples routinely analysed for microsporidian spores, only five were positive by microscopy. By comparison, 17 were positive for microsporidial DNA by real-time analysis, comprising 14 Enterocytozoon bieneusi, one Encephalitozoon cuniculi and two separate Pleistophora species infections.

scholarly journals Usefulness of Multiplex Real-Time PCR for Simultaneous Pathogen Detection and Resistance Profiling of Staphylococcal Bacteremia

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Yousun Chung ◽  
Taek Soo Kim ◽  
Young Gi Min ◽  
Yun Ji Hong ◽  
Jeong Su Park ◽  
...  
Keyword(s):  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
Pathogen Detection ◽  
Methicillin Resistance ◽  
Blood Cultures ◽  
Pcr Assay ◽  
Accurate Identification ◽  
Essential Information ◽  
Methicillin Resistant

Staphylococci are the leading cause of nosocomial blood stream infections. Fast and accurate identification of staphylococci and confirmation of their methicillin resistance are crucial for immediate treatment with effective antibiotics. A multiplex real-time PCR assay that targetsmecA,femAspecific forS. aureus,femAspecific forS. epidermidis,16S rRNAfor universal bacteria, and16S rRNAspecific for staphylococci was developed and evaluated with 290 clinical blood culture samples containing Gram-positive cocci in clusters (GPCC). For the 262 blood cultures identified to the species level with the MicroScan WalkAway system (Siemens Healthcare Diagnostics, USA), the direct real-time PCR assay of positive blood cultures showed very good agreement for the categorization of staphylococci into methicillin-resistantS. aureus(MRSA), methicillin-susceptibleS. aureus(MSSA), methicillin-resistantS. epidermidis(MRSE), methicillin-susceptibleS. epidermidis(MSSE), methicillin-resistant non-S. epidermidisCoNS (MRCoNS), and methicillin-susceptible non-S. epidermidisCoNS (MSCoNS) (κ=0.9313). The direct multiplex real-time PCR assay of positive blood cultures containing GPCC can provide essential information at the critical point of infection with a turnaround time of no more than 4 h. Further studies should evaluate the clinical outcome of using this rapid real-time PCR assay in glycopeptide antibiotic therapy in clinical settings.

A fluorescence-based quantitative real-time PCR assay for accurate Pocillopora damicornis species identification

Coral Reefs ◽  
2016 ◽  
Vol 35 (3) ◽  
pp. 895-899 ◽  
Author(s):  
Luke Thomas ◽  
Michael Stat ◽  
Richard D. Evans ◽  
W. Jason Kennington
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Pocillopora Damicornis ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay

Efficient species identification of pine marten (Martes martes) and red fox (Vulpes vulpes) scats using a 5′ nuclease real-time PCR assay

2007 ◽  
Vol 9 (3) ◽  
pp. 735-738 ◽  
Author(s):  
Catherine O’Reilly ◽  
Mark Statham ◽  
Jacinta Mullins ◽  
Peter D. Turner ◽  
Declan O’Mahony
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Vulpes Vulpes ◽  
Red Fox ◽  
Martes Martes ◽  
Pine Marten ◽  
Pcr Assay

scholarly journals Simple Real-Time PCR and Amplicon Sequencing Method for Identification of Plasmodium Species in Human Whole Blood

2015 ◽  
Vol 53 (7) ◽  
pp. 2251-2257 ◽  
Author(s):  
Martina I. Lefterova ◽  
Indre Budvytiene ◽  
Johanna Sandlund ◽  
Anna Färnert ◽  
Niaz Banaei
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Whole Blood ◽  
18S Rrna ◽  
Pcr Assay ◽  
Test Species ◽  
Content Type ◽  
The Real ◽  
Two Samples

Malaria is the leading identifiable cause of fever in returning travelers. AccuratePlasmodiumspecies identification has therapy implications forP. vivaxandP. ovale, which have dormant liver stages requiring primaquine. Compared to microscopy, nucleic acid tests have improved specificity for species identification and higher sensitivity for mixed infections. Here, we describe a SYBR green-based real-time PCR assay forPlasmodiumspecies identification from whole blood, which uses a panel of reactions to detect species-specific non-18S rRNA gene targets. A pan-Plasmodium18S rRNA target is also amplified to allow species identification or confirmation by sequencing if necessary. An evaluation of assay accuracy, performed on 76 clinical samples (56 positives using thin smear microscopy as the reference method and 20 negatives), demonstrated clinical sensitivities of 95.2% forP. falciparum(20/21 positives detected) and 100% for thePlasmodiumgenus (52/52),P. vivax(20/20),P. ovale(9/9), andP. malariae(6/6). The sensitivity of theP. knowlesi-specific PCR was evaluated using spiked whole blood samples (100% [10/10 detected]). The specificities of the real-time PCR primers were 94.2% forP. vivax(49/52) and 100% forP. falciparum(51/51),P. ovale(62/62),P. malariae(69/69), andP. knowlesi(52/52). Thirty-three specimens were used to test species identification by sequencing the pan-Plasmodium18S rRNA PCR product, with correct identification in all cases. The real-time PCR assay also identified two samples with mixedP. falciparumandP. ovaleinfection, which was confirmed by sequencing. The assay described here can be integrated into a malaria testing algorithm in low-prevalence areas, allowing definitivePlasmodiumspecies identification shortly after malaria diagnosis by microscopy.

scholarly journals Evaluation of High Resolution Melting (HRM) Analysis for Meat Species Identification of Raw and Cooked Meat

Separations ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 116
Author(s):  
Peyman Gholamnezhad ◽  
Hamed Ahari ◽  
Gholamreza Nikbakht Brujeni ◽  
Seyed Amir Ali Anvar ◽  
Abbasali Motallebi
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Meat Products ◽  
Pcr Assay ◽  
Hrm Analysis ◽  
Mitochondrial Cytochrome B ◽  
Meat Samples

The current study aimed to examine a real-time PCR assay with high-resolution melting (HRM) analysis for the species identification of minced meat samples. Meat samples from several animal species were purchased and minced separately or as a mixture of two species. DNA was extracted from all meat samples and subjected to real-time PCR assay by amplifying species-specific mitochondrial cytochrome b regions. Regarding the meat mixtures, two separate melting curves with specific melt peak temperatures (Tm) were detected. Additionally, DNA from each species was quantified, based on the calibration curves. The results showed that a real-time PCR assay with HRM analysis is suitable for the species identification of meat products, and could be used for the detection of meat frauds.

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