Evaluation of the Human Fragile X Mental Retardation 1 Polymerase Chain Reaction Reagents to Amplify theFMR1Gene: Testing in a Clinical Diagnostic Laboratory

Brucellosis is a widespread zoonosis. Currently the diagnosis of this zoonosis is based on microbiological and serological laboratory tests. Polymerase chain reaction (PCR) has been used to detect DNA from Brucella . Different target genes, primer pairs, PCR techniques, and extraction procedures have previously been published for Brucella detection. But only a few of these primers have been used in human samples, and only one study has been carried out to compare sensitivity between them. In the present study, 3 sets of primers and 3 different PCR protocols amplifying 3 different regions of the Brucella genome were compared for detection of Brucella DNA in a peripheral-blood PCR assay to conclude which is most suitable for the clinical diagnostic laboratory. These 3 pairs of primers amplify 3 different fragments included in (i) a gene encoding a 31 kDa Brucella abortus antigen (B4/B5), (ii) a sequence 16S rRNA of B. abortus (F4/R2), and (iii) a gene encoding an outer membrane protein (omp-2) (JPF/JPR). Some modifications on the reported techniques were applied during the present work to improve the outcome. The results showed that the B4/B5 primer pair had the highest sensitivity for detection of positive samples (98%), the JPF/JPR primer pair detected 88.4% of positive samples, whereas F4/R2 primer pair was the least sensitive, being able to detect only 53.1% of positive samples. The specificity of the 3 techniques was 100%. The B4/B5 primer pair was also able to detect the smallest number of bacteria (700 cfu/mL), whereas JPF/JPR was able to detect 7 × 105 cfu/mL and F4/R2 was able to detect 7 × 107cfu/mL. It is thus concluded that using the B4/B5 primer PCR with the suggested modifications is a robust assay, which meets the sensitivity requirements to be used for testing of human blood samples for brucellosis in the diagnostic laboratory.

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A Novel Polymorphism in the FMR1 Gene: Implications for Clinical Testing of Fragile X Syndrome

Archives of Pathology & Laboratory Medicine ◽

10.5858/2008-132-95-anpitf ◽

2008 ◽

Vol 132 (1) ◽

pp. 95-98

Author(s):

Bharat Thyagarajan ◽

Matthew Bower ◽

Michael Berger ◽

Sidney Jones ◽

Michelle Dolan ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Mental Retardation ◽

Fragile X Syndrome ◽

Southern Blot ◽

Trinucleotide Repeat ◽

Fragile X ◽

Fmr1 Gene ◽

Chain Reaction ◽

Cgg Repeat ◽

Polymerase Chain

Abstract Fragile X syndrome is the most common cause of inherited mental retardation among males. In most cases, the molecular basis of fragile X syndrome is the expansion and subsequent methylation of a CGG trinucleotide repeat in the 5′ untranslated region of the fragile X mental retardation 1 (FMR1) gene. Laboratory diagnosis usually relies on a combination of Southern blot and polymerase chain reaction analyses. In this case report we describe an unusual Southern blot result in a patient who presented with developmental delay and had a normal CGG repeat number by polymerase chain reaction analysis. Further investigation revealed a novel G3310C transversion in the FMR1 gene resulting in a new recognition site for the BssHII restriction enzyme. This novel restriction site could potentially mimic a partial deletion of the FMR1 gene on Southern blot analysis and thus represents a possible pitfall in the diagnosis of fragile X syndrome.

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Diagnostic validity of the chemiluminescent method compared to polymerase chain reaction for hepatitis B virus detection in the routine clinical diagnostic laboratory

Advanced Biomedical Research ◽

10.4103/2277-9175.133178 ◽

2014 ◽

Vol 3 (1) ◽

pp. 116 ◽

Cited By ~ 5

Author(s):

Yousef Rasmi ◽

Arsalan Ghavam ◽

Mohammad-Hassan Khadem-Ansari ◽

Mir-Davood Omrani

Keyword(s):

Polymerase Chain Reaction ◽

Hepatitis B Virus ◽

Virus Detection ◽

Diagnostic Validity ◽

Chain Reaction ◽

Clinical Diagnostic Laboratory ◽

Diagnostic Laboratory ◽

B Virus ◽

Polymerase Chain ◽

Chemiluminescent Method

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Puces à ADN comme outil d'identification moléculaire pour Culicoides

Revue d’élevage et de médecine vétérinaire des pays tropicaux ◽

10.19182/remvt.10054 ◽

2009 ◽

Vol 62 (2-4) ◽

pp. 144

Author(s):

J.C. De Witte ◽

I. Deblauwe ◽

Gill De Deken ◽

R. De Deken ◽

M. Madder ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Internal Transcribed Spacer ◽

Culicoides Species ◽

Chain Reaction ◽

Diagnostic Laboratory ◽

Laboratory Equipment ◽

Naked Eye ◽

Culicoides Obsoletus ◽

Polymerase Chain ◽

Obsoletus Complex

A DNA microarray test based on internal transcribed spacer 1 (ITS1) genotype expression was developed to identify Culicoides species of Northwestern Europe belonging to the Culicoides Obsoletus complex. The assay was designed so as to allow interpretation by the naked eye. False positive and false negative results were eliminated. The need for expensive laboratory equipment and reagents was kept as low as possible, making the technique affordable and feasible for any diagnostic laboratory with polymerase chain reaction (PCR) facilities. The microarray test could be validated through the three ringtests organised by Medreonet. Use of this microarray can improve monitoring adult and immature Culicoides species.

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