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. 2013 Jul;7(2):119-24.
doi: 10.4103/0973-6247.115568.

RHD alleles in the Tunisian population

Mouna Ouchari  1 , Saloua Jemni-Yaacoub, Taher Chakroun, Saida Abdelkefi, Batoul Houissa, Slama Hmida
Affiliations

RHD alleles in the Tunisian population

"V体育官网" Mouna Ouchari et al. Asian J Transfus Sci. 2013 Jul.

Abstract (V体育官网)

Background: A comprehensive survey of RHD alleles in Tunisia population was lacking. The aim of this study was to use a multiplex RHD typing assay for simultaneous detection of partial D especially with RHD/RHCE deoxyribonucleic acid (DNA) sequence exchange mechanism and some weak D alleles. VSports手机版.

Materials and methods: Six RHD specific primer sets were designed to amplify RHD exons 3, 4, 5, 6, 7 and 9 V体育安卓版. DNA from 2000 blood donors (1777 D+ and 223 D-) from several regions was selected for RHD genotyping using a PCR multiplex assay. Further molecular investigations were done to characterize the RHD variants that were identified by the PCR multiplex assay. .

Results: In the 1777 D+ samples, only 10 individuals showed the absence of amplification of exons 4 and 5 that were subsequently identified by PCR-SSP as weak D type 4 variants V体育ios版. No hybrid allele was detected. In the 223 D-, RHD amplification of some exons was observed only in 5 samples: 4 individuals expressed only RHD exon 9, and one subject lacking exons 4 and 5. These samples were then screened by PCR-SSPs on d(C) ce(s) and weak D type 4, respectively. .

Conclusion: The weak D type 4 appears to be the most common D variant allele. We have not found any partial D variant. Findings also indicated that RHD gene deletion is the most prevalent cause of the D- phenotype in the Tunisian population VSports最新版本. .

Keywords: Partial D; RHD genotyping; Tunisia; polymerase chain reaction multiplex; polymerase chain reaction with sequence-specific priming; weak D. V体育平台登录.

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"VSports最新版本" Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1
Representative results of a multiplex PCR analysis in our study. A 3% agarose gel of the PCR products obtained with genomic DNAs from a D+sample, a D− sample resulting from RHD gene deletion and different RHD variants. Lane 1: DNA marker; lane 2: Weak D type 4.0; lane 3: D+; lane 4: Variant (RHD exons 4 and 5 negatives); lane 5: Variant (only RHD exon 9+); lane 6, D (RHD exons 4 and 5 negatives); lane 7: D . C: Human growth hormone gene, which generating a 429 pb fragment, was used at internel control
Figure 2
Figure 2
Results of molecular RHD characterization. A schematic representation of the RHD typing strategy introduced in our blood transfusion center
Figure 3
Figure 3
(a) Results of PCR-SSP specific for T201V (C602G) substitution. (b) Results of PCR-SSP specific for T223V (T667G) substitution. Lane 1: Subject with weak D type 4.0 phenotype; lane 2: D+; lane 3: Variant carrying C602G and T667G substitutions. The upper bands indicate a 429-bp internal control amplicon which is positive in all cases. The lower bands correspond to the RHD specific product. M: molecular marker
Figure 4
Figure 4
(a) PCR-SSP specific for D-CE exon 3 hybrid: Lanes 1, 2 and 3: D+samples, Lanes 4, 5, and 6: 3 samples display the 110 pb product corresponding to the D-CE hybrid exon 3. (b) PCR-SSP specific for RHCE exon 5 polymorphism: Lane 1: Subject with RH: 34 (733G/733G), lane 2: hemizygous (733C/733G); lane 3 and 4: Homozygous (733C/733C). (c) PCR-SSP specific for RHCE exon 7 polymorphism: Lane 1: Subject with RH: 34 (1006T/1006T); lane 2: Hemizygous (1006G/1006T); lane 3: Homozygous (1006G/1006G). M: molecular marker
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