Recently-developed molecular assay loop-mediated isothermal amplification (LAMP) was also performed for further sensitive detection of parasite DNA.
Briefly, 10 ng RNA was reverse transcribed to cDNA with the Taqman microRNA Reverse Transcription kit and the Taqman microRNA assay stem loop primers (Applied Biosystems).
Briefly, RNA from 100 μl of serum was reverse transcribed to cDNA with the Taqman microRNA Reverse Transcription kit and the Taqman microRNA assay stem loop primers (Applied Biosystems).
For the LAMP assay with loop primers, amplification had been initially detected at 30 min, and reached the maximal at 45 min.
The assay of loop-mediated isothermal amplification (LAMP) is used to amplify target DNA from multidrug-resistant M. tuberculosis isolates, and enzyme-linked immunosorbent assay (ELISA) is used for the colorimetric determination.
In the rabbit ileal loop assay, where 104 wild type bacteria were sufficient to induce fluid accumulation, 109 Iexpressingessing CTS106 bacteria were needed to induce some fluid accumulation, while IEM101 expressing CTK63 was inactive, even when 1010 cells were used.
Results of the ileal loop assay suggest that the main influence is expressed at a stage beyond invasion.
Keleti & Sykora [ 169] also used a rabbit isolated ileal loop assay, which did not show any positive findings from the three cyanobacterial LPS isolates injected.
The invasion of mutant and wild type strains of G9 in the intestine of 10 12 week-old hens was investigated by an intestinal loop assay, as previously described [ 12].
Although basal levels of apoptosis were observed once epithelial cells underwent anoikis, a concentration- and time-dependent induction of cell death was observed in cells exposed in vivo (ligated loop assay) or in vitro to rGal-1.
For the displacement loop assay, the 5′ Cy3-labeled telomeric 90-mer [2.4 μM in nucleotides (nt), 26.67 nM oligo] was incubated with no protein or 1000 1500 nM Rad51 at 37 °C for 10 min in a reaction buffer containing 5 mM HEPES-KOH (pH 7.5), 1 mM CaCl, 1 mM ATP, 0.8 mM DTT, and 100 μg/mL BSA.
