Noninvasive Fetal Sex Determination by Real-Time PCR and TaqMan Probes

Ahmadi, Mohammad Hossein; Amirizadeh, Naser; Rabiei, Maryam; Rahimi-Sharbaf, Fatemeh; Pourfathollah, Ali Akbar

doi:10.29252/rbmb.9.3.315

Cited by 4 publications

(4 citation statements)

References 25 publications

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“…The percentage of fetal free DNA detected by ddPCR using RASSF1 and ACTB probes was 2%. 25 Figure 3 shows the detection results of ddPCR. Table 3 shows 26 blood samples collected from pregnant women of different gestational ages (weeks 5–39) and the predicted and actual blood groups of the fetus, which all predicted prenatal blood groups that were confirmed after birth.…”

Section: Resultsmentioning

confidence: 99%

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Non-invasive prenatal testing for fetal Ss, Kidd, and CTL2 blood group prediction by multiplex digital droplet PCR

Wang

Chu

Chen

et al. 2023

Therapeutic Advances in Hematology

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Background: Some blood groups, such as S and s blood groups in the MNS blood group system, and Kidd and CTL2 blood group systems, can cause severe fetal and newborn alloimmune disorders. Non-invasive prenatal testing (NIPT) to predict fetal blood groups and knowledge of local blood group gene frequency are both important for pregnancy management decisions. Droplet digital PCR (ddPCR) has high specificity and sensitivity in detecting fetal single nucleotide variation. Objectives: The objective is to predict fetal Ss, Kidd, and CTL2 blood groups using multiplex ddPCR. The gene frequencies of three blood groups were detected by ddPCR in northwest China. Design: This is a prospective study. Methods: Cell-free fetal DNA isolated from 26 healthy single pregnant women at different gestational stages was tested with QX200 Droplet Digital PCR. Results were compared with fetal genotypes. DNA samples purified from 20 blood pools containing a total of 1000 donors in northwest China were subjected to ddPCR to detect the gene frequency of three blood groups. Results: Ss, Kidd, and CTL2 blood groups of 26 pregnant fetuses were accurately detected by multiplex ddPCR. The multiplex ddPCR results were consistent with the Sanger sequencing results of 26 fetal blood samples after birth. The gene frequencies of the three blood groups detected by ddPCR were 9.30% for S, 90.70% for s, 48.43% for Jka, 51.57% for Jkb, 66.57% for HNA-3A, and 33.43% for HNA-3B. Conclusions: It is reliable to predict fetal Ss, Kidd, and CTL2 blood groups by multiplex ddPCR. Meanwhile, we designed a simple and efficient method for inferring the gene frequency of three blood groups based on ddPCR.

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Section: Resultsmentioning

confidence: 99%

“…Conversely, if the fetus is homozygous, an allele imbalance occurs. By using SRY and RASSF1 genes, 25 which indicate fetal gene content, we can determine fetal genotypes.…”

Section: Nipt Of Fetal Blood Groups Using Ddpcr Assaysmentioning

confidence: 99%

Non-invasive prenatal testing for fetal Ss, Kidd, and CTL2 blood group prediction by multiplex digital droplet PCR

Wang

Chu

Chen

et al. 2023

Therapeutic Advances in Hematology

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“…Under appropriate conditions, the enzyme cleans CGCG sites from guanine (G) to cytosine (C). Moreover, the enzyme is sensitive to methylation and cannot recognize methylated CGCG sites [ 27 , 28 ]. The specificity of BstUI restriction endonuclease site recognition was used to digest the nucleic acid, and specific primers were designed for fluorescence quantitative PCR detection to improve detection sensitivity.…”

Section: Discussionmentioning

confidence: 99%

Multiple Biomarker-Combined Screening for Colorectal Cancer Based on Bisulfate Conversion-Free Detection of Fecal DNA Methylation

Liu¹,

Liu

et al. 2021

BioMed Research International

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To evaluate the applicability of bisulfate conversion-free methylation assay based on enzyme digestion in fecal screening for colorectal cancer (CRC). Stool samples were collected from a total of 1142 participants with intestinal abnormalities, including 180 positive cases, 60 advanced adenomas, and 902 negative cases. DNA from reference cell lines and clinical samples was extracted and digested with an enzyme to detect the methylation of CRC markers SEPT9, SDC2, NDRG4, SFRP2, and BMP3 genes. Statistical analysis was then used to determine the ability of the markers, both individually and in combination, to detect CRC and adenoma. Our results showed that the enzyme digestion method could suitably detect DNA marker methylation in as low as 1% of the cell lines. BMP3 had a considerably low detection rate in all clinical samples, with only 6 positive cases detected out of 180 cancer samples. Our findings showed that the combination of SEPT9, SDC2, and SFRP2 had an area under the receiver operation curve of 0.937, sensitivity of 94.11%, and specificity of 89.21% for detecting CRC. Moreover, the detection sensitivity of adenoma can also reach 38.33%. After innovatively utilizing bisulfate conversion-free methylation assay for CRC screening, this study verified the potential clinical applicability of combining multiple biomarkers for CRC screening in a large number of samples.

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“…In the past, researchers have used several tools to study the genetic properties of T. vaginalis, including Restriction Fragment Length Polymorphism (RFLP) (13), internal transcribed spacer analysis (ITS) (14), quantitative real-time PCR (15) and nucleotide sequencing (PCR sequencing) (16). Interestingly, the actin gene was found to induce morphological changes and play a major role in the pathogenesis of T. vaginalis (17) making it an ideal candidate for targeted investigations on the molecular level, and genotyping (including PCR sequencing) (18).…”

Section: Introductionmentioning

confidence: 99%

Identification of Trichomonas Vaginalis Genotypes Using by Actin Gene and Molecular Based Methods in Southwest of Iran

et al. 2021

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Background:Trichomonas vaginalis (T. vaginalis) is a sexually transmitted protozoan parasite and the causative agent of trichomoniasis. The genetic characterization of T. vaginalis isolates shows notable genetic variation in this parasite. In the present study, we aimed to identify the T. vaginalis genotypes based on analyzing of actin gene in women specimens referred to health centers of Ilam city, southwest Iran. Methods: A total of 1765 female samples were collected from gynecology clinics in the city of Ilam. DNA was extracted from positive samples and nested polymerase chain reaction (Nested PCR) was used to amplify the actin gene. Then, partial sequencing and genotyping of the actin gene was performed. A phylogenetic tree was drawn using the detected genotypes of T. vaginalis and reference sequences. Results: Twenty-one of the 1765 urine and vaginal samples were positive for T. vaginalis. All infected individuals were married and their age in years was between 25 to 34. Further, the majority of infected women had cervical lesions, patchy erythema, and white color discharge. According to sequencing analysis, the isolates were identified as genotype G (n= 8) and genotype E (n= 2). Conclusions: From the collected samples, we were able to distinguish at least two genotypes (G and E) of T. vaginalis. However, lesser is known about these genotypes in the city of Ilam. Further studies with a higher number of isolates should be performed in order to understand the implications of these results in this region.

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Noninvasive Fetal Sex Determination by Real-Time PCR and TaqMan Probes

Cited by 4 publications

References 25 publications

Non-invasive prenatal testing for fetal Ss, Kidd, and CTL2 blood group prediction by multiplex digital droplet PCR

Non-invasive prenatal testing for fetal Ss, Kidd, and CTL2 blood group prediction by multiplex digital droplet PCR

Multiple Biomarker-Combined Screening for Colorectal Cancer Based on Bisulfate Conversion-Free Detection of Fecal DNA Methylation

Identification of Trichomonas Vaginalis Genotypes Using by Actin Gene and Molecular Based Methods in Southwest of Iran

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