scholarly journals Molecular Markers for Human Sex Determination in Forensic Genetics Analysis

2021 ◽  
Vol 8 (6) ◽  
pp. 25-30
Author(s):  
Maan H. Salih
Keyword(s):  
Sex Determination ◽  
Multiplex Pcr ◽  
Genetic Marker ◽  
Y Chromosome ◽  
Internal Control ◽  
Forensic Genetics ◽  
Sex Identification ◽  
Pcr Analysis ◽  
Positive Band ◽  
Two Samples

Sex determination is indispensable in forensic anthropology, sexual disorder, and also as part of large-scale genetic population studies. The purpose of this investigation is to determine the human sex from whole blood using multiplex PCR analysis. Blood samples from 75 male and 70 female healthy volunteers were taken from Tikrit city, Iraq. Our study identified a reliable set of three primer locus, namely SRY, ALT1 (internal control) and amelogenin locus. The SRY primer on the Y chromosome showed a 254 bp of PCR product, with 100% accuracy for human male identification. Thus, the pair of SRY primers was considered a strong genetic marker for human sex identification. Amelogenin regions in the Y chromosome showed a true positive band (236 bp) with 100% accuracy on sex identification. Amelogenin regions in X chromosome also showed positive bands (330 bp) in female samples and positive band in male samples except for two samples showed a negative band (null bands). The most obvious finding from this study is that multiplex PCR of ALT1 and SRY is consider as a reliable genetic marker for human sex identification. The research has also shown that amelogenin is good genetic marker for human sex identification.

369 AMPLIFICATION OF THE SRY GENE FOR SEXING OF BUFFALO (BUBALUS BUBALIS) EMBRYOS USING NESTED MULTIPLEX PCR

2007 ◽  
Vol 19 (1) ◽  
pp. 300
Author(s):  
M. Zhang ◽  
Q. Fu ◽  
W. S. Qin ◽  
H. Y. Zheng ◽  
Y. Q. Lu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Internal Control ◽  
Pcr Amplification ◽  
Single Copy ◽  
Bubalus Bubalis ◽  
Sex Identification ◽  
Sry Gene ◽  
Embryo Sex ◽  
Pcr Method ◽  
Male Specific

In mammals, the Y chromosome-linked SRY gene is responsible for male sex determination. Therefore, a logical approach for embryo sex identification is to amplify the male-specific single-copy SRY gene. The objective of this study was to design specific primers for amplification of buffalo SRY gene and develop a reliable PCR method for sex identification of buffalo embryos. Genomic DNA was extracted from swamp buffalo (Bubalus bubalis) peripheral blood. A pair of primers based on the sequence of Holstein bovine SRY gene (forward, 52-GTTTGCCTTATGGATTTATT-32; reverse, 52-TCTACTTTAGCCTATTTG-32) was used to amplify whole buffalo SRY gene. This amplified fragment was isolated and constructed into plasmids for sequencing. Two pairs of primers, S1/S2 (forward, 52-CCATGAACGCCTTCATTTTGTG-32; reverse, 52-ACGAGGTCGATATTTATAGC CC-32) and S3/S4 (forward, 52-AAGCAGCTGGGCTATGAGTGGAA-32; reverse, 52-ACGAGGTCGATATTTATAGCCC-32), were designed based on the SRY sequence above. Simultaneously, the G3PDH gene was amplified to serve as an internal control for both male and female embryos. A multiplex-nested-PCR system was optimized by varying the following parameters individually: concentration of Mg2+, dNTPs, primers, and different cycles number. Twenty-seven IVF morulae were identified with the optimal PCR procedure after biopsy. Accuracy of PCR amplification was verified by dot blotting. The sex of 24 embryos fertilized with Y-sperm separated by flow cytometry was also examined. Results indicated that the optimal procedure of Nested-Multiplex-PCR consisted of 1.5 mM Mg2+, 100 �M dNTPs, 0.5 �M SA3/SA4 primers, and 0.25 �M GA3/GA4 primers, and 35 cycles. Accuracy of identification was 100% for 27 IVF morulae; 14 were judged as males and 13 were females. The result of blotting confirmed that the accuracy of amplification was 100%. The proportion of males was 83.3% (20/24) in embryos fertilized with Y-sperm. This confirms that the PCR system targeting the SRY gene can be used for accurate sex identification of buffalo embryos. This study was supported by grants from the Foundation of Guangxi Key Laboratory for Subtropical Bio-Resource Conservation and Utilization (SB0403) and the Guangxi Department of Science and Technology (0626001-3-1).

scholarly journals Study of Y-Chromosome Microdeletions in Azoospermic Infertile Males using Multiplex PCR Analysis

2018 ◽  
Vol 15 (2) ◽  
pp. 351-357 ◽  
Author(s):  
Prafulla S. Ambulkar ◽  
Sunil S. Pande
Keyword(s):  
Multiplex Pcr ◽  
Y Chromosome ◽  
Pcr Analysis ◽  
Obstructive Azoospermia ◽  
Time Saving ◽  
Pcr Screening ◽  
Azoospermia Factor ◽  
Sts Markers ◽  
Y Chromosome Microdeletions ◽  
Severe Impairment

The infertility affects about 15% of couples and male factors being responsible about 40-50%. In male infertility, genetic abnormalities of Y chromosome play crucial role in spermatogenesis defect. Y chromosome q arm having Azoospermia factor region (AZFa, AZFb, and AZFc) are most important for spermatogenesis. Here, we investigated the frequencies of Y-chromosome microdeletions using three sets of multiplex PCR in idiopathic cases of azoospermia. We studied a total of 110 infertile male with non-obstructive azoospermia subjects & 50 fertile control subjects. All DNA samples were used for Y chromosome microdeletions analysis by using 11 STS markers in three different multiplex PCR of AZF regions. Out of 110 infertile azoospermic males, 14 (12.72%) infertile males showed partial deletion of AZF regions using three sets of multiplex PCR group. In the AZF microdeletions of infertile males, individually AZFc region was the most deletions sites (10%) followed by AZFb (6.36%) and AZFa (1.81%). The sites and sizes of microdeletions differ in all infertile azoospermic males who showed at least two or more STS markers microdeletions. The frequency of Y chromosome microdeletions in our azoospermic infertile males is 12.72%. We conclude that Y chromosome microdeletions frequency in azoospermic infertile males is higher than other infertile group due to severe impairment in spermatogenesis. Multiplex PCR screening of microdeletions is very useful and time saving technique when used more number of STS markers. It will be great help to infertility clinics for genetic counseling and assisted reproduction.

scholarly journals Evaluation of an internal control in a multiplex-PCR assay for sex determination of <i>in vitro</i>-produced bovine embryos

2011 ◽  
Vol 02 (06) ◽  
pp. 456-462
Author(s):  
Fabiana de Almeida Rufino ◽  
Marcelo Marcondes Seneda ◽  
Alice Fernandes Alfieri ◽  
Katia Cristina Silva-Santos ◽  
Karina Cristina Puggesi Rubin ◽  
...  
Keyword(s):  
Sex Determination ◽  
Multiplex Pcr ◽  
Internal Control ◽  
Bovine Embryos ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Spot Urine for Sex Determination Forensic Identifications with Amelogenin Locus and Y chromosomes (DYS 19). Bercak Urin untuk Identifikasi Forensik Jenis Kelamin dengan Lokus Amelogenin dan Y Kromosom (DYS19)

2018 ◽  
Vol 20 (3) ◽  
pp. 180
Author(s):  
Yeti Eka Sispita Sari
Keyword(s):  
Sex Determination ◽  
Y Chromosome ◽  
X Chromosome ◽  
Sex Chromosome ◽  
Dna Amplification ◽  
Single Copy ◽  
Amelogenin Gene ◽  
Spot Urine ◽  
Y Chromosomes ◽  
Two Samples

AbstractBackground:  Amelogenin gene was a single copy gene located in an X chromosome and a Y chromosome. The location of amelogenin gene for identification of sex chromosome has good variability between the form and the shape of the X chromosome and the Y chromosome and between Amelogenin alleles among different populations. Purpose: To prove urine spot examination on the results of the sex determination through Deoxyribo Nucleid Acid (DNA) isolation using amelogenin and Y chromosome loci (DYS19). Methods: Spotting the microscopic examination of urine samples to determine the presence or absence of urethral epithelial cells, followed by isolation Deoxyribo nucleid Acid (DNA) in order to determine the extent and purity of DNA amplification. Then performed Polymerase Chain Reaction (PCR) amelogenin locus at 106bp - 112bp and Y chromosomes (DYS19) at 232 -268 bp. Results: in 9 samples of men from 3 families with 3 kinship of different regions shows the results of different tests, because Amel Y variation between individual and populations method of determining the sex of 100% was inaccurate. In some men Amel Y can be removed entirely. This research should be visualized one band on the Y chromosome (DYS19) and the Amelogenin two bands during electrophoresis occurs misidentification of the sample as a woman. Conclusions: Identification of sex using Amelogenin locus and Y chromosomes (DYS19) has six identical and ambiguous results because the two samples shown as the sign of men but visualized as women, another sample was not visualized because of the thick level and concentration of Deoxyribo nucleid Acid (DNA).Keywords: Urine Spot, Sex Determination, Amelogenin, Y chromosome (DYS19).

scholarly journals MULTIPLEX-PCR FOR DETECTION OF β-LACTAM RESISTANCE IN Staphylococcus spp.

2019 ◽  
Vol 6 (2) ◽  
pp. 262-275
Author(s):  
Giovana Hashimoto Nakadomari ◽  
Amanda Carmen Charalo ◽  
Ana Claudia Lemes Pavan ◽  
Vanessa Kelly Capoia Vignoto ◽  
Ricardo Antonio Pilegi Sfaciotte ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Resistance Genes ◽  
Internal Control ◽  
Meca Gene ◽  
Simultaneous Detection ◽  
Rrna Gene ◽  
16S Gene ◽  
Internal Reaction ◽  
Two Samples ◽  
Staphylococcus Spp

A Staphylococcus Multiplex PCR system was developed for the simultaneous detection of the mecA, mecC, blaZ (resistance genes of b-lactam resistance) and PVL (pathogenicity factor gene), associated with an internal reaction control with the 16S rRNA gene. There were used primers described in the literature with and without modification and designed primers to standardize the hybridization and amplification temperature of distinct bands with 139 bp (mecC), 228 bp (16S), 313 bp (mecA), 408 bp (PVL) and 516 bp (blaZ) of molecular weight. The standardization was performed in ATCC strains and Staphylococcus schleiferiand tested in 56 strains of Staphylococcusspp. The 16S gene (internal control) was amplified in all samples, mecA gene was detected in two samples, mecA associated with mecC gene in one sample, mecA associated to the blaZ gene in 14 samples and the blaZ gene in 15 samples. No resistance genes were amplified in 24 samples. The PVL gene was not amplified in any of the samples tested.

Sex determination and Y chromosome constitutive heterochromatin

2019 ◽  
Vol 1 (1) ◽  
pp. 1-5
Author(s):  
Abyt Ibraimov
Keyword(s):  
Sex Determination ◽  
Y Chromosome ◽  
Constitutive Heterochromatin ◽  
Sex Differentiation ◽  
Secondary Sexual Characteristics ◽  
Biological Meaning ◽  
Heterochromatin Block ◽  
Sexual Characteristics ◽  
Open Question ◽  
Efficient Elimination

In many animals, including us, the genetic sex is determined at fertilization by sex chromosomes. Seemingly, the sex determination (SD) in human and animals is determined by the amount of constitutive heterochromatin on Y chromosome via cell thermoregulation. It is assumed the medulla and cortex tissue cells in the undifferentiated embryonic gonads (UEG) differ in vulnerability to the increase of the intracellular temperature. If the amount of the Y chromosome constitutive heterochromatin is enough for efficient elimination of heat difference between the nucleus and cytoplasm in rapidly growing UEG cells the medulla tissue survives. Otherwise it doomed to degeneration and a cortex tissue will remain in the UEG. Regardless of whether our assumption is true or not, it remains an open question why on Y chromosome there is a large constitutive heterochromatin block? What is its biological meaning? Does it relate to sex determination, sex differentiation and development of secondary sexual characteristics? If so, what is its mechanism: chemical or physical? There is no scientifically sound answer to these questions.

scholarly journals Seroepidemiological study of toxoplasmosis in women referred to a pre-marriage counseling center in Alborz Province, Iran

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Melica Shahighi ◽  
Aliehsan Heidari ◽  
Hossein Keshavarz ◽  
Amir Bairami ◽  
Saeedeh Shojaee ◽  
...  
Keyword(s):  
Congenital Toxoplasmosis ◽  
Pcr Analysis ◽  
Women Of Childbearing Age ◽  
Childbearing Age ◽  
Serum Samples ◽  
Chi Square ◽  
Igg Avidity ◽  
Two Samples ◽  
Chi Square Test ◽  
Associated Risk Factors

Abstract Objectives The aim of the current study was to assess prevalence of Toxoplasma infection and its associated risk factors in women of childbearing-age in central Iran. Results Of 400 serum samples assessed for anti-T. gondii antibodies, 81 (20.25%) samples were positive for anti-T. gondii antibodies, including 74 positive samples (91.3%) for anti-T. gondii IgG and seven positive samples (8.7%) for IgG and IgM. Of seven IgG and IgM positive samples, five and two samples were high and low in IgG avidity, respectively. Based on PCR analysis, Toxoplasma infection was detected in one sample with anti-T. gondii IgM and low IgG avidity. The Chi-square test showed significant correlations of T. gondii seropositivity with history of undercooked meat consumption and contacts with cats (p < 0.05). In the present study, 79.75% of the participants were negative for IgG against T. gondii infection. Furthermore, recently acquired Toxoplasma infection was found using IgG avidity and PCR assays among women of childbearing-age in the study area, which would increase the risk of their fetus becoming infected. Educational program and antenatal screening of childbearing-age women for T. gondii infection may be important primary prevention strategies and help reduce the risk of congenital toxoplasmosis in this population.

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