scholarly journals Improved Protocol for Using Avian Red Blood Cells as Substrates for the Polymerase Chain Reaction

BioTechniques ◽  
1999 ◽  
Vol 26 (6) ◽  
pp. 1080-1082 ◽  
Author(s):  
Dani Bercovich ◽  
Yoram Plotsky ◽  
Yosef Gruenbaum
Keyword(s):  
Polymerase Chain Reaction ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals DETEKSI HEMAGLUTININ, HEMOLISIN DAN KOAGULASE SECARA FENOTIPIK DAN GENOTIPIK PADA Staphylococcus aureus ISOLAT ASAL BROILER

2018 ◽  
Vol 36 (1) ◽  
pp. 103
Author(s):  
Khusnan Khusnan ◽  
Dwi Kusmanto
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Wound Infections ◽  
Rabbit Plasma ◽  
Chain Reaction ◽  
Sheep Red Blood Cells ◽  
Polymerase Chain

Staphylococcus aureus is a bacterium causing diseases in animals and human. Staphylococcus aureus in broilers cause septicemia, tendosinovitis, dermatitis, endocarditis, wound infections, arthritis and bumblefoot. In this research, 24 isolates of Staphylococcus aureus from broiler were characterized of its virulent factors including the presence of haemagglutinin, ability to agulate the plasma in tubular coagulase test as well presipitate formation in clumping factor test, and haemolysis types. By polymerase chain reaction (PCR) were genotypically detected genes coa, clf, hlaA, and hlaB. All of the isolates (100%) had haemagglutinin, capable to agglutinate and precipitae of rabbit plasma. All isolates could lyse sheep red blood cells with the type of α-hemolysis (45.8%),  β-hemolysis (50.0%) and γ-haemolysis (4.2%). Genotypically, all isolates (100%) had coa and clf genes,  hlaA gene 7(0.8%) and hlaB gene (29.2%).

scholarly journals Red blood cells in cerebrospinal fluid as possible inhibitory factor for enterovirus RT-PCR

2016 ◽  
Vol 74 (10) ◽  
pp. 810-815 ◽  
Author(s):  
Sérgio Monteiro de Almeida ◽  
Sônia Mara Raboni ◽  
Meri Bordignon Nogueira ◽  
Luine R. Renaud Vidal
Keyword(s):  
Polymerase Chain Reaction ◽  
Cerebrospinal Fluid ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Virus Detection ◽  
Inhibitory Factor ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Negative Results ◽  
Polymerase Chain

ABSTRACT The presence of hemoglobin in samples are considered an important inhibitory factor for polymerase chain reaction (PCR). The aim of this study was to examine the influence of red blood cells (RBC)s in cerebrospinal fluid (CSF) as an inhibitory factor to reverse transcription polymerase chain reaction (RT-PCR) for enteroviruses (EV). Forty-four CSF samples from patients showing characteristics of viral meningitis were assessed for EV by RT-PCR. Viral RNA extracted with guanidine isothyocianate buffer and virus detection was performed by in-house nested PCR. Positivity for EV RT-PCR was higher in CSF samples without RBCs than in samples with RBCs: 13(26%) and 36(9.2%), p = 0.001. In the group with positive EV RT-PCR, the mean + SD CSF RBC was 37 ± 183 cell/mm3; the group with negative results had 580 + 2,890 cell/mm3 (p = 0.007). The acceptable upper limit for CSF RBCs that could not influence RT-PCR was 108 cells/mm3. CSF samples with negative results for EV RT-PCR have more erythrocytes.

scholarly journals Detection of the molecular abnormality in chronic myeloid leukemia by use of the polymerase chain reaction

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 2063-2065
Author(s):  
A Dobrovic ◽  
KJ Trainor ◽  
AA Morley
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Myeloid Leukemia ◽  
Blood Cells ◽  
Myeloid Leukemia ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cell ◽  
Great Sensitivity ◽  
Chain Reaction ◽  
Molecular Abnormality ◽  
Polymerase Chain

The bcr-abl translocation characteristic of chronic myeloid leukemia (CML) was detected by the polymerase chain reaction (PCR) modified to use mRNA as the starting material. Amplification of a sequence spanning the bcr-abl junction was obtained by using peripheral blood cells from all of 20 patients with classic CML, one patient with acute lymphoblastic leukemia probably secondary to CML, and two cell lines derived from patients with CML. The presence of bcr exon 3 in the mRNA was determined from the size of the amplified sequence; it was present in 14 and absent in seven patients. One leukemic cell per 1,000 nonleukemic cells could be readily detected, thus indicating the great sensitivity of the method. This technique is of routine value in CML both for diagnosis and for following the course of treatment.

scholarly journals Detection of residual leukemia after bone marrow transplant for chronic myeloid leukemia: role of polymerase chain reaction in predicting relapse

Blood ◽  
1991 ◽  
Vol 77 (4) ◽  
pp. 874-878 ◽  
Author(s):  
TP Hughes ◽  
GJ Morgan ◽  
P Martiat ◽  
JM Goldman
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Chronic Myeloid Leukemia ◽  
Bone Marrow Transplant ◽  
Blood Cells ◽  
Myeloid Leukemia ◽  
Marrow Transplant ◽  
Early Assessment ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract We used the polymerase chain reaction (PCR) to detect residual leukemia- specific mRNA in blood and marrow from 37 patients in complete hematologic and cytogenetic remission after allogeneic bone marrow transplant (BMT) for chronic myeloid leukemia (CML). Our two-step PCR method involved the use of “nested primers” in the second step and could detect one K562 cell diluted into 10(5) normal cells. Elaborate measures were taken to exclude false-positive and false-negative results. In nine patients whose blood and marrow were studied simultaneously the results were concordant (two positive and seven negative). Twenty-three patients transplanted in chronic phase (CP) with unmanipulated donor marrow were studied. Blood cells from nine of these patients were studied 3 to 6 months post-BMT and six were PCR positive; three were negative on subsequent studies. Blood cells from 18 patients studied between 8 months and 8 years post-BMT were all PCR negative. Nine patients transplanted in CP with T-cell-depleted marrow cells were studied. Blood from five was positive 3 to 24 months post- BMT; blood from five was negative 3 to 6 years post-BMT. Four patients no longer in first CP were studied after BMT with unmanipulated donor marrow. Blood from all four was positive 5 to 19 months post-BMT. Based on the known clinical results of transplant in these three cohorts we conclude that PCR may be positive within 6 months of BMT in patients who can expect long-lasting remission, whereas PCR positivity later after BMT may indicate that the probability of cure is reduced. Thus, the technique may prove useful for early assessment of new transplant protocols that might inadvertently increase the risk of relapse.

scholarly journals New sensitive method for the detection of the A3243G mutation of human mitochondrial deoxyribonucleic acid in diabetes mellitus patients by ligation-mediated polymerase chain reaction

1998 ◽  
Vol 44 (10) ◽  
pp. 2088-2093 ◽  
Author(s):  
Michiyo Urata ◽  
Machiko Wakiyama ◽  
Masanori Iwase ◽  
Makoto Yoneda ◽  
Sachiko Kinoshita ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Polymerase Chain Reaction ◽  
Conventional Method ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Nucleotide Position ◽  
Peripheral Blood Cells ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sensitive Method

Abstract An adenine-to-guanine mutation at nucleotide position (np) 3243 in the mitochondrial tRNALeu(UUR) gene is closely associated with various clinical phenotypes of diabetes mellitus. Because the mutation creates a new restriction site for the restriction enzyme ApaI, the mutation is usually detected and quantified by ApaI cleavage of the PCR products including np 3243. The sensitivity of the conventional method is, however, 5–10% heteroplasmy. The percentage of heteroplasmy of the mutation is usually highest in the affected tissues and is much lower in peripheral blood cells, which are used most frequently for the analysis. The sensitivity of the conventional method, however, is not sufficient to detect the mutation from peripheral blood cells. Utilizing ligation-mediated polymerase chain reaction, we have developed a feasible and sensitive method to detect 0.01% heteroplasmy of the 3243 mutation in peripheral leukocytes.

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