Simultaneous differential detection of human pathogenic and nonpathogenicVibriospecies using a multiplex PCR based ongyrB andpntA genes

Teh, Cindy Shuan Ju; Chua, Kek Heng; Thong, Kwai Lin

doi:10.1111/j.1365-2672.2009.04599.x

Cited by 30 publications

(29 citation statements)

References 17 publications

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“…In silico PCR as described previously was carried out prior to actual PCR evaluation to make sure the primers used are targeted to the correct loci (Teh et al, 2010;Thong et al, 2011). The forward and reverse primers are 5ꞌ-CTG GAG ACC ACT CCC ATC CTT TCT-3ꞌ and 5ꞌ-GAT GTG GCC ATC ACA TTC GTC AGA T-3ꞌ, respectively.…”

Section: Methodsmentioning

confidence: 99%

Angiotensin-converting enzyme gene I/D dimorphism does not play a major role in the susceptibility of Malaysian systemic lupus erythematosus patients

Lian¹,

Lau²,

Ching³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Systemic lupus erythematosus (SLE) is an autoimmune disease that causes systemic damage, involving auto-reactive antibodies and over-deposition of immune complexes. Susceptibility to SLE is believed to be multifactorial, and genetics is one of the proven etiological factors; it can affect SLE development, severity and prognosis. We investigated a possible association between the angiotensin-converting enzyme gene and susceptibility to SLE in the Malaysian population. PCR was employed for the determination of I/D dimorphism of this gene. The I allele was more frequent than the D allele in both the SLE patients (N = 170) and healthy controls (N = 190). However, there was no significant difference in the distribution of these two alleles between both groups studied (χ 2 = 0.284, P > 0.05). Interestingly, the DD homozygous genotype scored notably higher in the healthy control group (χ 2 = 7.568, P < 0.05), while the ID heterozygote was observed to be significantly associated with SLE (χ 2 = 11.143, P < 0.05). In conclusion, with respect to the Malaysian population, the DD genotype might play a protective role in the development of SLE while in contrast, those who carry the ID genotype might be at potential risk for onset of this disease.

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Section: Methodsmentioning

confidence: 99%

Angiotensin-converting enzyme gene I/D dimorphism does not play a major role in the susceptibility of Malaysian systemic lupus erythematosus patients

Lian¹,

Lau²,

Ching³

et al. 2012

Genet. Mol. Res.

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“…Furthermore, PCR facilitates the identification of the strains that are viable but non-culturable [12] in this study 16S rRNA gene was amplified and showed great results. It is less labor intensive and much faster than conventional methods, and that is the reason of its increasing application among investigators [13]. The antibiogram profile revealed that isolates, all isolates showed multi-drug resistant to amoxicillin and nitrofurantoin.…”

Section: Discussionmentioning

confidence: 99%

Antibiogram and Molecular Characterization of Vibrio Cholerae Isolated From Marine Fish

Angelo

Ramesh

2017

Asian J Pharm Clin Res

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Objective: Molecular identification and antibiotic susceptibility evaluation of Vibrio cholerae from marine fish available in local fish market Thanjavur, Tamil Nadu, India.Methods: Inoculation was done using nutrient agar as general media and thiosulfate-citrate-bile salts-sucrose agar as selective media and confirmed as V. cholerae by Gram-stain (microscopic observation), growth characteristics of different media, biochemical tests such as methyl red test, nitrate reduction tests, and indole test. Sensitivity (drug sensitivity) was done in Mueller-Hinton agar using disc diffusion method 10 different antibiotics were used to evaluate the antibiogram profile, molecular detection was done by targeting 16S rRNA gene using a universal primer.Results: V. cholerae is present in marine fish samples, as showed by culture method and microscopic observation as well biochemical tests. Polymerase chain reaction (PCR) amplification of 16S rRNA gene showed the amplification of targeted gene and antibiogram profile showed that isolates are more sensitive to ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V. cholerae infection by the physicians and amoxicillin must be avoided which is resistant. Conclusion:Molecular detection is safe and rapid methods for bacteria identification as revealed by PCR amplification of 16S rRNA gene. As the isolates are more sensitive to ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V. cholerae infection by the physicians and amoxicillin, and nitrofurantoin must be avoided.

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“…Currently, multiplex PCR is widely applied for the rapid diagnosis of many infectious diseases (20,31,32,34). With well-designed primers used under fully optimized conditions, most of the multiplex systems have shown high sensitivity and specificity.…”

Section: Discussionmentioning

confidence: 99%

“…The primers with the secondary structure were redesigned with slight modifications. The selected primers were then tested through a BLAST search and in silico PCR (http: //insilico.ehu.es/PCR/) as described previously for the prediction of amplicons (32,34). PCR conditions for each of the primer pairs were optimized individually and then mixed together to perform a multiplex PCR for the detection of five species of human malaria parasites.…”

Section: Microscopy Examinationmentioning

confidence: 99%

Hexaplex PCR Detection System for Identification of Five Human Plasmodium Species with an Internal Control

et al. 2012

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Malaria remains one of the major killers of humankind and persists to threaten the lives of more than one-third of the world's population. Given that human malaria can now be caused by five species of Plasmodium, i.e., Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, and the recently included Plasmodium knowlesi, there is a critical need not only to augment global health efforts in malaria control but also, more importantly, to develop a rapid, accurate, species-sensitive/species-specific, and economically effective diagnostic method for malaria caused by these five species. Therefore, in the present study, a straightforward single-step hexaplex PCR system targeting five human Plasmodium 18S small-subunit rRNAs (ssu rRNAs) was designed, and the system successfully detected all five human malaria parasites. In addition, this system enables the differentiation of single infection as well as mixed infections up to the two-species level. This assay was validated with 50 randomly blinded test and 184 clinical samples suspected to indicate malaria. This hexaplex PCR system is not only an ideal alternative for routine malaria diagnosis in laboratories with conventional PCR machines but also adds value to diagnoses when there is a lack of an experienced microscopist or/and when the parasite morphology is confusing. Indeed, this system will definitely enhance the accuracy and accelerate the speed in the diagnosis of malaria, as well as improve the efficacy of malaria treatment and control, in addition to providing reliable data from epidemiological surveillance studies.

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Simultaneous differential detection of human pathogenic and nonpathogenicVibriospecies using a multiplex PCR based ongyrB andpntA genes

Cited by 30 publications

References 17 publications

Angiotensin-converting enzyme gene I/D dimorphism does not play a major role in the susceptibility of Malaysian systemic lupus erythematosus patients

Angiotensin-converting enzyme gene I/D dimorphism does not play a major role in the susceptibility of Malaysian systemic lupus erythematosus patients

Antibiogram and Molecular Characterization of Vibrio Cholerae Isolated From Marine Fish

Hexaplex PCR Detection System for Identification of Five Human Plasmodium Species with an Internal Control

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