Real-Time PCR Approaches for Analysis of Hydrocarbon-Degrading Bacterial Communities

2015 ◽  
pp. 45-64 ◽  
Author(s):  
Boyd A. McKew ◽  
Cindy J. Smith
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Communities

scholarly journals Lack of Evidence for Microbiota in the Placental and Fetal Tissues of Rhesus Macaques

mSphere ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Kevin R. Theis ◽  
Roberto Romero ◽  
Andrew D. Winters ◽  
Alan H. Jobe ◽  
Nardhy Gomez-Lopez
Keyword(s):  
Dna Sequencing ◽  
Microbial Communities ◽  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Communities ◽  
Rhesus Macaques ◽  
Uterine Wall ◽  
Bacterial Burden ◽  
Quantitative Real Time Pcr ◽  
Multiple Modes

ABSTRACT The prevailing paradigm in obstetrics has been the sterile womb hypothesis. However, some are asserting that the placenta, intra-amniotic environment, and fetus harbor microbial communities. The objective of this study was to determine whether the fetal and placental tissues of rhesus macaques harbor bacterial communities. Fetal, placental, and uterine wall samples were obtained from cesarean deliveries without labor (∼130/166 days gestation). The presence of bacteria in the fetal intestine and placenta was investigated through culture. The bacterial burden and profiles of the placenta, umbilical cord, and fetal brain, heart, liver, and colon were determined through quantitative real-time PCR and DNA sequencing. These data were compared with those of the uterine wall as well as to negative and positive technical controls. Bacterial cultures of fetal and placental tissues yielded only a single colony of Cutibacterium acnes. This bacterium was detected at a low relative abundance (0.02%) in the 16S rRNA gene profile of the villous tree sample from which it was cultured, yet it was also identified in 12/29 background technical controls. The bacterial burden and profiles of fetal and placental tissues did not exceed or differ from those of background technical controls. By contrast, the bacterial burden and profiles of positive controls exceeded and differed from those of background controls. Among the macaque samples, distinct microbial signals were limited to the uterine wall. Therefore, using multiple modes of microbiologic inquiry, there was not consistent evidence of bacterial communities in the fetal and placental tissues of rhesus macaques. IMPORTANCE Microbial invasion of the amniotic cavity (i.e., intra-amniotic infection) has been causally linked to pregnancy complications, especially preterm birth. Therefore, if the placenta and the fetus are typically populated by low-biomass microbial communities, current understanding of the role of microbes in reproduction and pregnancy outcomes will need to be fundamentally reconsidered. Could these communities be of benefit by competitively excluding potential pathogens or priming the fetal immune system for the microbial bombardment it will experience upon delivery? If so, what properties (e.g., microbial load and community membership) of these microbial communities preclude versus promote intra-amniotic infection? Given the ramifications of the in utero colonization hypothesis, critical evaluation is required. In this study, using multiple modes of microbiologic inquiry (i.e., culture, quantitative real-time PCR [qPCR], and DNA sequencing) and controlling for potential background DNA contamination, we did not find consistent evidence for microbial communities in the placental and fetal tissues of rhesus macaques.

scholarly journals Dynamics of Vaginal Bacterial Communities in Women Developing Bacterial Vaginosis, Candidiasis, or No Infection, Analyzed by PCR-Denaturing Gradient Gel Electrophoresis and Real-Time PCR

2007 ◽  
Vol 73 (18) ◽  
pp. 5731-5741 ◽  
Author(s):  
Beatrice Vitali ◽  
Ciro Pugliese ◽  
Elena Biagi ◽  
Marco Candela ◽  
Silvia Turroni ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Relative Abundance ◽  
Real Time Pcr ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Pcr Analysis ◽  
Healthy Women ◽  
Gradient Gel Electrophoresis

ABSTRACT The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.

DGGE and real-time PCR analysis of lactic acid bacteria in bacterial communities of the phyllosphere of lettuce

2008 ◽  
Vol 52 (5) ◽  
pp. 614-623 ◽  
Author(s):  
Jutta Zwielehner ◽  
Michael Handschur ◽  
Astrid Michaelsen ◽  
Selen Irez ◽  
Michael Demel ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Communities ◽  
Pcr Analysis

Feldvalidierung einer real-time PCR zum Nachweis von Mycoplasma hyopneumoniae in Nasentupfermaterial von lebenden Schweinen

2005 ◽  
Vol 147 (9) ◽  
pp. 373-379 ◽  
Author(s):  
F. Zeeh ◽  
P. Kuhnert ◽  
R. Miserez ◽  
M. G. Doherr ◽  
W. Zimmermann
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Mycoplasma Hyopneumoniae

Nutzen des hoch-sensitiven real-time-PCR HBV Tests (TaqMan) zur Prädiktion der Resistenzentwicklung bei Lamivudin-Langzeittherapie

2010 ◽  
Vol 48 (08) ◽  
Author(s):  
A Brodzinski ◽  
F van Bömmel ◽  
B Fülöp ◽  
B Schlosser ◽  
M Biermer ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr
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