Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification - Fig 2

<p>(a) Effective capture of primer on supplimer of sPIN particle. In order to see the capturing efficiency, sPIN particles were prepared with matched (red) or mismatched (black) supplimer to primer. After incubation with FAM-modified primer, both of them showed high intensity depending on the injected FAM-primer concentration (filled red and black circle). After rinsing, mismatched sPIN particle lost the fluorescence (empty black circle), meanwhile matched sPIN particle showed consistently remained fluorescence (empty red circle). It was caused by the stable binding between the primer and corresponding supplimer. (b) Positive qPCR graph of each sPIN particle using supplimers with different T_m. (c) Negative qPCR graph of each sPIN particle with supplimers of different T_m. (d) Fluorescent signal subtracted No Template Control (NTC) signal from Positive Template Control (PTC) showing regular S-shaped curves.</p

PCR reaction with sPIN particle and multiplex qPCR.

<p>PCR reaction with sPIN particle and multiplex qPCR.</p

Integral optical density of Calprotectin and TLR4 protein expression in jejunal mucosa.

a, b<p>Means with different superscripts in the same row differ (<i>P</i><0.05).</p>1<p>A group means the control group, B group means the chlortetracycline group, C group means the COS group.</p

Calprotectin and TLR4 protein expression in jejunal mucosa (immunohistochemical staining, ×400).

<p>Piglets challenged by enterotoxigenic <i>Escherichia</i> coli were fed either a control diet (A group, n = 6), or control diet plus chlortetracycline (B group, n = 6) or control diet plus chitosan (C group, n = 6) for 21 days and 6 pigs per treatment were killed to excise the jejunum.</p

Changes in diarrhea index in piglets.

<p>Piglets challenged by enterotoxigenic <i>Escherichia</i> coli were fed either a control diet (A group, n = 10), or control diet plus chlortetracycline (B group, n = 10) or control diet plus chitosan (C group, n = 10) for 21 days. Diarrhea index (%) was calculated as 100× number of piglets that had diarrhea/total number of piglets.</p

Serum IL-1β, IL-6 and TNF-α levels in weaned piglets.

1<p>A group means the control group, B group means the chlortetracycline group, C group means the COS group.</p

Sequences (5′–3′) of the primers used for the detection of mRNA specific for IL-1β, IL-6, TNF-α and GAPDH.

<p>Notes: F = forward primer; R = reverse primer.</p

Composition and nutrient levels of the basal diet (DM basis) %.

1<p>Provided additional vitamins per kilogram diet: VA 11 000 IU, VD₃ 1 100 IU, VE 16 IU, VK 1 mg, pantothenate 6 mg, retinoic acid 2 mg, folic acid 0.8 mg, nicotinic acid 10 mg, thiamine 0.6 mg, VB₁ 0.6 mg, biotin 0.08 mg, VB₁₂ 0.03 mg.</p>2<p>Provided with additional trace elements per kilogram diet: Zn 165 mg, Fe 165 mg, Mn 33 mg, Cu 16.5 mg, I 297 µg, Se 297 µg.</p

The relative expression of IL-1β, IL-6 and TNF-α mRNA in jejunal mucosa.

<p>Piglets challenged by enterotoxigenic <i>Escherichia</i> coli were fed either a control diet (A group, n = 6), or control diet plus chlortetracycline (B group, n = 6) or control diet plus chitosan (C group, n = 6) for 21 days and 6 pigs per treatment were killed to excise the jejunum.</p