PRIMARY SLX TEST USING REAL-TIME PCR BASED ON HIGH RESOLUTION MELTING (HRM) ON MICROFILARIA EXAMINATION

Background: Filariasis patients can be a source of transmission if their blood still contains microfilariae. One of the Polymerase Chain Reaction (PCR) methods used is High Resolution Melting (HRM), using primary specificity testing. Purpose: To test the specificity of SLX primer. The samples used for this test were isolates of Salmonella., Klebsiella, Pseudomonas, negative and positive controls for Brugia malayi and Wuchereria bancrofti. Method: The design in this study is a quasi-experiment by testing the specificity of SLX primer using HRMbased real-time PCR based on the Cycle Threshold (CT) value observed through the amplification curve. Result: The real-time PCR results showed that no CT was released in the bacterial samples, and there was a CT value in the positive control. The results of this study indicate that specific SLX primer can be used in identifying microfilariae. Conclusion: SLX primer have a reasonable specificity because they cannot detect the existence of microorganisms in the samples other than microfilariae.

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The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and discrimination of nematodes Bursaphelenchus xylophilus and Bursaphelenchus mucronatus

Molecular and Cellular Probes ◽

10.1016/j.mcp.2016.02.003 ◽

2016 ◽

Vol 30 (2) ◽

pp. 113-117 ◽

Cited By ~ 6

Author(s):

Anna Filipiak ◽

Beata Hasiów-Jaroszewska

Keyword(s):

Polymerase Chain Reaction ◽

High Resolution ◽

Real Time ◽

Real Time Pcr ◽

High Resolution Melting ◽

Bursaphelenchus Xylophilus ◽

Chain Reaction ◽

Hrm Analysis ◽

Bursaphelenchus Mucronatus ◽

Polymerase Chain

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The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and identification of pathogenetic and nonpathogenetic populations of the quarantine nematode Bursaphelenchus xylophilus

Progress in Plant Protection ◽

10.14199/ppp-2017-034 ◽

2017 ◽

Vol 57 (3) ◽

Keyword(s):

Polymerase Chain Reaction ◽

High Resolution ◽

Real Time ◽

Real Time Pcr ◽

High Resolution Melting ◽

Bursaphelenchus Xylophilus ◽

Chain Reaction ◽

Hrm Analysis ◽

Detection And Identification ◽

Polymerase Chain

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Complement C3 Genotyping of Slow and Fast Variants by Real Time PCR-High Resolution Melting

European Journal of Inflammation ◽

10.1177/1721727x1201000308 ◽

2012 ◽

Vol 10 (3) ◽

pp. 329-334 ◽

Cited By ~ 2

Author(s):

D.M. Valero-Hervás ◽

P. Morales ◽

M.J. Castro ◽

P. Varela ◽

M. Castillo-Rama ◽

...

Keyword(s):

High Resolution ◽

Real Time ◽

Real Time Pcr ◽

High Resolution Melting ◽

Low Cost ◽

Complement C3 ◽

Amplification Refractory Mutation System ◽

Rt Pcr ◽

Dna Amount ◽

Mutation System

“Slow” and “Fast” C3 complement variants (C3S and C3F) result from a g.304C>G polymorphism that changes arginine to glycine at position 102. C3 variants are associated with complement-mediated diseases and outcome in transplantation. In this work C3 genotyping is achieved by a Real Time PCR - High Resolution Melting (RT-PCR-HRM) optimized method. In an analysis of 49 subjects, 10.2% were C3FF, 36.7% were C3SF and 53.1% were C3SS. Allelic frequencies (70% for C3S and 30% for C3F) were in Hardy-Weinberg equilibrium and similar to those published previously. When comparing RT-PCR-HRM with the currently used Tetraprimer-Amplification Refractory Mutation System PCR (T-ARMS-PCR), coincidence was 93.8%. The procedure shown here includes a single primer pair and low DNA amount per reaction. Detection of C3 variants by RT-PCR-HRM is accurate, easy, fast and low cost, and it may be the method of choice for C3 genotyping.

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