scholarly journals Detection of Methanobrevobacter smithii and Methanobrevibacter oralis in Lower Respiratory Tract Microbiota

2020 ◽  
Vol 8 (12) ◽  
pp. 1866
Author(s):  
Yasmine Hassani ◽  
Fabienne Brégeon ◽  
Gérard Aboudharam ◽  
Michel Drancourt ◽  
Ghiles Grine

Methanogens, the sole microbes producing methane, are archaea commonly found in human anaerobic microbiota. Methanogens are emerging as opportunistic pathogens associated with dysbiosis and are also detected and cultured in anaerobic abscesses. Their presence in the respiratory tract is yet unknown. As a preliminary answer, prospective investigation of 908 respiratory tract samples using polyphasic approach combining PCR-sequencing, real-time PCR, fluorescent in situ hybridization (FISH), and methanogens culture was carried out. Methanobrevibacter smithii and Methanobrevibacter oralis DNA sequences, were detected in 21/527 (3.9%) sputum samples, 2/188 (1.06%) bronchoalveolar lavages, and none of 193 tracheo-bronchial aspirations. Further, fluorescence in situ hybridization detected methanogens in three sputum investigated specimens with stick morphology suggesting M. oralis and in another one bronchoalveolar lavage sample investigated, diplococal morphology suggesting M. smithii. These observations extend the known territory of methanogens to the respiratory tract and lay the foundations for further interpretation of their detection as pathogens in any future cases of isolation from bronchoalveolar lavages and the lungs.

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 640-643 ◽  
Author(s):  
N Cuñado ◽  
E Sánchez-Morán ◽  
J Barrios ◽  
J L Santos

Some Alliaceae species have no tandemly repeated TTTAGGG sequences. Instead, at the very end of their chromosomes, there are highly repetitive satellite and (or) rDNA sequences. These sequences apparently replace the canonical plant telomeric sequences in these species. A method of preparing two-dimensional surface spreads of plant synaptonemal complexes (SCs), combined with fluorescent in situ hybridization, has revealed that telomeric chromatin is tightly condensed at the ends of SCs in plants and animals. Using this method, we have tested the organization and location of those sequences postulated to cap the chromosomes in two species of the genus Allium: A. cepa and A. altaicum. We have also extended this study to other putative telomere candidates, such as LTR (long terminal repeat) and non-LTR retrotransposons. None of the DNA sequences analyzed showed the characteristic telomeric organization at pachytene.Key words: fluorescent in situ hybridization, meiosis, repetitive DNA, Allium, synaptonemal complex.


Genome ◽  
2000 ◽  
Vol 43 (1) ◽  
pp. 181-184 ◽  
Author(s):  
Chunxian Chen ◽  
Huihuang Yan ◽  
Wenxue Zhai ◽  
Lihuang Zhu ◽  
Jingsan Sun

Two clones of a new family of tandemly repeated DNA sequences have been isolated from a maize random genomic DNA library. MR68 is 410 bp, representing a monomeric unit and MR77 is 1222 bp, containing three units. The copy number was estimated to be about 3000 per 1C maize genome. Its methylation pattern was also determined. Fluorescent in situ hybridization (FISH) indicates that the sequence is located on the subtelomeric region of the long arm of chromosomes 3 and 6, as well as on the satellite of chromosome 6. Key words: Zea mays, tandemly repeated DNA, satellite DNA, fluorescent in situ hybridization (FISH).


2000 ◽  
Vol 118 (4) ◽  
pp. A50 ◽  
Author(s):  
Luigi Ricciardiello ◽  
Ajay Goel ◽  
Dong K. Chang ◽  
Christina L. Chang ◽  
C. Richard Boland

Genome ◽  
1995 ◽  
Vol 38 (4) ◽  
pp. 795-802 ◽  
Author(s):  
Angeles Cuadrado ◽  
Nicolás Jouve

The molecular characterization of C-banded regions of Secale montanum Guss. by means of in situ hybridization was performed in order to provide new information about their chromosome structure relative to cultivated rye, Secale cereale L. Accurate identification of individual chromosomes was achieved using simultaneous and (or) successive fluorescent in situ hybridization (FISH) and C-banding. FISH identification was performed using total rye DNA, three highly repetitive rye DNA sequences (pSc119.2, pSc74, and pSc34), and the ribosomal RNA probes pTa71 (18S, 5.8S, and 26S rDNA) and pTa794 (5S rDNA). FISH was also used to identify the chromosome segment involved in two spontaneous translocation lines recovered from a 'Chinese Spring' – S. montanum wheat–rye addition line. FISH analysis revealed the exact translocation breakpoints and allowed the identification of the transferred rye segments. The value of this type of analysis is discussed.Key words: Secale cereale, Secale montanum, rye, repetitive DNA, fluorescence in situ hybridization.


Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.


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