Population dynamics and in situ kinetics of nitrifying bacteria in autotrophic nitrifying biofilms as determined by real-time quantitative PCR

2006 ◽  
Vol 94 (6) ◽  
pp. 1111-1121 ◽  
Author(s):  
Tomonori Kindaichi ◽  
Yoshiko Kawano ◽  
Tsukasa Ito ◽  
Hisashi Satoh ◽  
Satoshi Okabe
Keyword(s):  
Population Dynamics ◽  
Real Time ◽  
Quantitative Pcr ◽  
Nitrifying Bacteria ◽  
Real Time Quantitative Pcr ◽  
Kinetics Of

scholarly journals Assessment of the Kinetics of Treponema pallidum Dissemination into Blood and Tissues in Experimental Syphilis by Real-Time Quantitative PCR

2007 ◽  
Vol 75 (6) ◽  
pp. 2954-2958 ◽  
Author(s):  
Juan C. Salazar ◽  
Asha Rathi ◽  
Nelson L. Michael ◽  
Justin D. Radolf ◽  
Linda L. Jagodzinski
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Whole Blood ◽  
Mononuclear Cells ◽  
Treponema Pallidum ◽  
Blood Components ◽  
Peripheral Blood Mononuclear ◽  
Real Time Quantitative Pcr ◽  
Kinetics Of

ABSTRACT Little is known about the size and kinetics of treponemal burdens in blood and tissues during acquired or experimental syphilitic infection. We used real-time quantitative PCR to measure Treponema pallidum DNA levels in rabbits infected intratesticularly with the prototype Nichols strain. At the outset, we performed a series of in vitro blood spiking experiments to determine the effect of blood processing procedures on the distribution of treponemes in various blood components. T. pallidum DNA levels in plasma and whole blood were approximately 10-fold higher than those in serum and more than 200-fold greater than those in peripheral blood mononuclear cells (PBMCs). Ten rabbits were inoculated intratesticularly with doses of treponemes ranging from 4 × 107 to 2 × 108 organisms. In five rabbits, T. pallidum DNA levels were measured sequentially in serum, plasma, whole blood, and PBMCs until sacrifice at peak orchitis, at which time brain, kidney, liver, spleen, and testicles were harvested; blood and organs were also harvested at orchitis from the other five rabbits. T. pallidum DNA was detected in plasma within 24 h postinfection. Treponeme levels in whole blood and blood components increased significantly with the development of peak orchitis. Overall, levels in serum and PBMCs were lower than those in plasma and whole blood; this disparity was particularly marked at early time points. Significantly greater numbers of spirochetes were found in the spleen than in liver, kidney, or brain tissue at the time of sacrifice. Our findings highlight the remarkable capacity of T. pallidum to disseminate from the site of infection to blood and tissues, and they identify the spleen as a prime target for treponemal invasion.

Kinetics of Early Donor Chimerism after Nonmyeloablative Haematopoietic Stem Cell Transplantation Monitored with High-Resolution Real Time Quantitative PCR.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3669-3669
Author(s):  
Tania N. Masmas ◽  
Soren L. Petersen ◽  
Hans O. Madsen ◽  
Lars Ryder ◽  
Ebbe Dickmeiss ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Resolution ◽  
Real Time ◽  
Quantitative Pcr ◽  
Haematopoietic Stem Cell ◽  
Logrank Test ◽  
Post Transplant ◽  
Real Time Quantitative Pcr ◽  
Donor Chimerism ◽  
Kinetics Of

Abstract Purpose: From May 2003 to March 2005, 37 patients with haematological malignancies were transplanted with peripheral blood stem cells after nonmyeloablative conditioning with fludarabine 30 mg/m2 i.v. once daily on day −4 to −2 and 200 cGY of total body irradiation on day 0. Post-transplant immunosuppression consisted of oral cyclosporin 12.5 mg/kg/day and mycophenolate mofetil 30 mg/kg/day. Three patients were not eligible for follow-up due to lack of informed consent or informative markers for chimerism measurement. The purpose of this study was to evaluate kinetics of early chimerism of CD4+, CD8+, and, CD15+ cells immediately post transplant and furthermore to identify factors associated with rejection of graft. Methods: Blood samples were collected post transplant on day +1–3, +5–7, and then weekly. Samples were separated with immunomagnetic beads and DNA was salt extracted. Chimerism analysis was performed by automated high-resolution real-time quantitative PCR based on short insertion or deletion polymorphisms or single nucleotide polymorphisms. Results: The kinetics of CD15+ chimerism differed from that of CD4+ and CD8+ chimerism (Figure). CD15+ donor chimerism was low with a median of 0.9% donor cells on day +1–3, and remained low until around day +14. Subsequently the donor chimerism increased rapidly towards 100%. The CD4+ and CD8+ donor chimerism in contrast started higher with a median of 16.3% and 15.1% respectively on day +1–3, increased slowly, and took months to reach 100%. CD4+, CD8+ or CD15+ donor chimerism on day +1–3 did not predict later development of acute graft versus host disease. Two patients had primary rejection of the donor stem cells, while four patients had secondary rejection. The patients with primary rejection had no measurable donor chimerism in any cell lineages from day +1–3. No patient with CD15+ donor chimerism above the median (0.9%) on day +1–3 rejected, while 40% (6/15) of the patients with CD15+ donor chimerism below the median rejected (LogRank test, p=0.01). No differences were found in total number of nucleated cells, CD34+, CD3+, CD4+, or CD8+ cells in the donor harvest between patient who rejected and those who did not. Conclusions: The kinetics of early CD15+ donor chimerism differed from the kinetics of early CD4+ and CD8+ donor chimerism. Early CD15+ donor chimerism was markedly lower than CD4+ and CD8+ donor chimerism. CD15+ donor chimerism increased toward 100% donor chimerism more rapidly than the CD4+ and CD8+ chimerism. Primary rejection of donor stem cells can be identified early, as these patients have no measurable donor CD4+ or CD8+ chimerism on day +1–3. Furthermore, patients with CD15+ donor chimerism above the median on day +1–3 have a low risk for rejection. Figure Figure

Detection of H19 expression in mouse placenta by real-time quantitative PCR and in situ hybridization

Placenta ◽  
2015 ◽  
Vol 36 (10) ◽  
pp. A6-A7
Author(s):  
Banyar Than Naing ◽  
Xiaohui Song ◽  
Toshihiro Takizawa
Keyword(s):  
In Situ Hybridization ◽  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Quantitative Pcr ◽  
Mouse Placenta

scholarly journals New Real-Time Quantitative PCR Procedure for Quantification of Bifidobacteria in Human Fecal Samples

2004 ◽  
Vol 70 (7) ◽  
pp. 4165-4169 ◽  
Author(s):  
Miguel Gueimonde ◽  
Satu Tölkkö ◽  
Teemu Korpimäki ◽  
Seppo Salminen
Keyword(s):  
In Situ Hybridization ◽  
Detection Limit ◽  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Pcr ◽  
Fluorescent In Situ Hybridization ◽  
The Real ◽  
Real Time Quantitative Pcr ◽  
Lanthanide Chelate

ABSTRACT The application of a real-time quantitative PCR method (5′ nuclease assay), based on the use of a probe labeled at its 5′ end with a stable, fluorescent lanthanide chelate, for the quantification of human fecal bifidobacteria was evaluated. The specificities of the primers and the primer-probe combination were evaluated by conventional PCR and real-time PCR, respectively. The results obtained by real-time PCR were compared with those obtained by fluorescent in situ hybridization, the current gold standard for intestinal microbiota quantification. In general, a good correlation between the two methods was observed. In order to determine the detection limit and the accuracy of the real-time PCR procedure, germfree rat feces were spiked with known amounts of bifidobacteria and analyzed by both methods. The detection limit of the method used in this study was found to be about 5 × 104 cells per g of feces. Both methods, real-time PCR and fluorescent in situ hybridization, led to an accurate quantification of the spiked samples with high levels of bifidobacteria, but real-time PCR was more accurate for samples with low levels. We conclude that the real-time PCR procedure described here is a specific, accurate, rapid, and easy method for the quantification of bifidobacteria in feces.

Replication kinetics of duck virus enteritis vaccine virus in ducklings immunized by the mucosal or systemic route using real-time quantitative PCR

2009 ◽  
Vol 86 (1) ◽  
pp. 63-67 ◽  
Author(s):  
Xuefeng Qi ◽  
Xiaoyan Yang ◽  
Anchun Cheng ◽  
Mingshu Wang ◽  
Yufei Guo ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Vaccine Virus ◽  
Real Time Quantitative Pcr ◽  
Kinetics Of ◽  
Duck Virus Enteritis
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