Combined use of multiplex ligation-dependent probe amplification and automatic sequencing for identification of KAL1 defects in patients with Kallmann syndrome.

[kallmann syndrome]

To investigate the role of KAL 1 abnormalities in Brazilian patients with Kallmann syndrome .In Â vitro experiments .Academic medical center .One hundred fifteen Brazilian patients (98 men ) with Kallmann syndrome .Peripheral blood leukocytes were used to obtain DNA .Direct sequencing and multiplex ligation-dependent probe amplification were used to identify KAL 1 abnormalities .We identified four KAL 1 mutations (p .Met 1 ?, p .Ala 33 Glyfs , p .Arg 257 *, and p .Trp 462 *) and two multiple exon deletions (exons 1 -2 and 3 -14 ) in six new male patients . Overall , 17 KAL 1 defects (14 .8 %) were identified in the entire cohort of patients with Kallmann syndrome , including previously studied cases . KAL 1 -mutated patients presented with a more severe reproductive and nonreproductive phenotype (synkinesia , renal malformations , cryptorchidism , and anatomic olfactory abnormalities ) in comparison with patients without KAL 1 mutations . Intragenic deletions were one of the most often encountered defects (29 .4 %). These Â deletions can be missed by polymerase chain reaction (PCR ) due to Yq 11 .2 KAL 1 pseudogene (KALP ) spurious amplification .These results indicate that intragenic multiexon deletions are one of the most frequent KAL 1 abnormalities , which can be more accurately detected by multiplex ligation-dependent probe amplification . In addition , KAL 1 sequencing results should be interpreted with caution , and stringency conditions of the PCR reaction should be adjusted to avoid pseudogene amplification .

Diseases
Validation

Diseases presenting "peripheral blood leukocytes" symptom

classical phenylketonuria

cowden syndrome

kallmann syndrome

von hippel-lindau disease

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