primary amenorrhea is the absence of menstruation and secondary sexual characteristics in phenotypic women aged 14 years or older .
common hormonal cause of primary amenorrhea includes pituitary dysfunction , chronic systemic disease , and absent ovarian function .
gonadal failure in genetically xx individuals is ovarian failure ; when this occurs at any time before onset of sexual maturation , there will be primary amenorrhea and incomplete breast development .
xy individuals with gonadal failure will have female genitalia because mullerian inhibiting factor and testosterone will not be produced .
gonadal tumors occur in up to 25% of women with a y chromosome ; unlike complete androgen insensitivity , these gonads do not secrete hormones and should be removed at the time of diagnosis .
the incidence of the chromosomal abnormality ( ca ) in live births is around 90 per 10,000 .
included in the incidence are the numerical ( monosomy / trisomy / mosaicism ) as well as the structural ( translocation / isochromosome / deletion / duplication / ring ) ca .
233 women with primary amenorrhea who were referred to cytogenetic unit , children hospital , mansoura university , egypt from july 2008 to december 2010 .
for all patients , steroidal hormonal assay were setup according to elisa technique,[810 ] and chromosomal cultures were setup according to g- banding , about 1 ml of blood was mixed with 5 ml of rpmi medium , 1 ml of fetal bovine serum , 0.1 g / ml of phytohemagglutinin ( pha ) and was incubated at 37c . after 72 hour of incubation , the colcimid ( 1 mg / ml ) was added and incubated for another 1.5 hour .
the cells were then harvested by hypotonic treatment ( 1.5 hour with 0.075 m kcl at 37c ) , fixed and washed thrice with fixative solution ( methanol and acetic acid in a ratio of 3:1 ) , and then metaphases were spread and stained using standard g - banding technique . for each case , 50 spread metaphases were analyzed with cytovision system .
fluorescence in situ hybridization ( fish ) technique was carried out in some of cases to more evaluation according to technique of .
10 l of the x , y centromeric probe were applied to the target area , and the cover slip was applied to the slide immediately .
the cover slip was sealed with rubber cement , and the slide was placed on hot plate at 72c for 2 minutes .
the slide was placed in a warmed humidified box , and the box was placed at 37c in an incubator overnight .
the cover slip was removed from slide , and it was immersed in 70 ml of the 0.4 x ssc/0.3% np-40 in a coplin jar at a 731c water bath for 2 minutes .
then , the slide was immersed in 70 ml of the 2 x ssc/0.1% np-40 in coplin jar at 25c for 2 minutes .
the slide was air - dried in the dark , and 10 l of dapi ii counter stain were applied to the target area of slide , and the cover slip was applied to the each slide .
233 patients with primary amenorrhea were referred from different parts of egypt to cytogenetic laboratory of genetic unit , children hospital mansoura university , from july 2008 to december 2010 .
( 1 ) the numerical abnormalities of the x chromosome were detected in 23 ( 50% ) .
( 2 ) structural abnormalities of the x chromosome were detected in 11 ( 23.91% ) .
( 4 ) male karyotype 46 , xy was presented in 2 ( 4.35% ) , [ table 1 ] .
the two patients with association of chromosome y was confirmed by fish [ figure 1 ] , and the presence of deletion of long arm of chromosome x in 3 patients was confirmed , using fish [ figure 2 ] . classification of chromosomal abnormalities of patients with primary amenorrhea interphase and metaphase from fish analysis showing one x chromosome and one y chromosome of female confirming the testicular feminization syndrome ( xy ) metaphase from fish analysis showing one x chromosome and short arm of another x of female confirming the form x , xq in table 2 , follicle stimulating hormone ( fsh ) and luteinizing hormone ( lh ) mean levels in sera of females with primary amenorrhea , whether they have numerical , structural , or mosaicism chromosomal abnormalities , were significantly higher than those of the control group ( p<0.05 ) .
but , estradiol ( e2 ) mean levels in sera of them were significantly lower than those of the control group ( p<0.05 ) .
20.63% of chromosomal abnormalities in primary amenorrhea of present study compared with those in various study conducted between 1983 and 2010 [ table 3 ] .
chromosome abnormalities in primary amenorrhea in various study and compared with present study genes essential for gonadal function are located on the proximal part of xp , the long arm of x proximal to xq13 and/or the long arm of x distal to xq26 .
large deletion of xq with breakpoints at or proximal to q13 are expected to produce gonadal dysgenesis with primary amenorrhea , half of the patients with such deletions have turner`s syndrome . that explained the cause of our two patients with deletion of xq have primary amenorrhea , but have not feature of turner syndrome , whereas other three patients who have deletion of xp have primary amenorrhea and all the features of turner syndrome .
ten et al . found one patient with primary amenorrhea and 47 , xxx had mental retardation and the cause of that , an extra x , can slow down embryonic cell development in a special way .
similar to that , there were mental retardation in two of our patients with karyotype of 47 , xxx .
xy individuals with gonadal failure will have female genitalia because mullerian inhibiting factor and testosterone will not be produced .
gonadal tumors occur in up to 25% of women with a y chromosome ; unlike complete androgen insensitivity , these gonads do not secrete hormones and should be removed at the time of diagnosis . in our study , we diagnosed two patients of primary amenorrhea as testicular feminization syndrome ( xy ) using fish technique . the molecular cytogenetic technique , fish , accurately delineate the nature and origin of marker chromosomes in primary amenorrhea patients , which is difficult by conventional cytogenetics .
as similar to that , in present study , the presence of deletion of long arm of chromosome x in 2 patients was confirmed , using fish , which is very difficult by conventional cytogenetic where short arm of the x chromosome is similar to the y chromosome .
ten et al . found that , serum follicle stimulating hormones and luteinizing hormones were always increased in primary amenorrhea patients with chromosomal abnormalities .
the present study showed that karyotype and fish are necessary to detect the causes of primary amenorrhea .
this study also revealed the incidence of chromosomal abnormalities in women with primary amenorrhea in egypt is similar to that reported in previous literatures .
 background : primary amenorrhea is defined as the absence of menstruation and secondary sexual characteristics in phenotypic women aged 14 years or older . 
 hormonal disorders are main causes of primary amenorrhea . 
 common hormonal cause of primary amenorrhea includes pituitary dysfunction and absent ovarian function . 
 the aim of this study was to estimate the incidence and types of chromosomal abnormalities in patients with primary amenorrhea in egypt.materials and methods : chromosomal analysis and hormonal assay were carried out on 223 patients with primary amenorrhea that were referred from different parts of egypt to cytogenetic laboratory of genetic unit , children hospital mansoura university , from july 2008 to december 2010 . 
 fish technique was carried out in some of cases to more evaluation.results:the frequency of chromosomal abnormalities was 46 ( 20.63% ) in primary amenorrhea patients . 
 the chromosomal abnormalities can be classified into four main types . 
 ( 1 ) the numerical abnormalities of the x  chromosome were detected in 23 ( 50 % ) . 
 ( 2 ) structural abnormalities of the x chromosome were detected in 11 ( 23.91% ) . 
 ( 3 ) mosaicism of x chromosome was found in 10 ( 21.74% ) . 
 ( 4 ) male karyotype 46 , xy was presented in 2 ( 4.35%).conclusion : the present study showed that karyotype and fish are necessary to detect the causes of primary amenorrhea . 
 this study also revealed the incidence of chromosomal abnormalities in women with primary amenorrhea in egypt is similar to that reported in previous literatures . 
Introduction
Materials and Methods
Results
Discussion
Conclusion