Tissue Typing Dept wrote:
>> For class I work I personally think that serology is the easiest and
> cheapest.
>> Derek O'Neill
> Tissue Typing Dept.
> Beaumont Hospital
> Dublin
>>tt at iol.ie>>Jonathan_Kurtis at Brown.edu (Jonathan Kurtis) wrote:
>> >Hi all,
>> >I will be doing some HLA class 1 typing of about 100 individuals from
> >Western Kenya. I will be using cryopreserved PBMCs as source material.
> >Does anyone have any thoughts as to which method is easier/better (PCR vs.
> >Ab+Complement). Are there other techniques?? Any advice on companies that
> >sell kits or usefull references?? I am particularly interested in the B
> >locus.
>> >Thanks for your help,
>> >jake
>> >--
> >Jonathan Kurtis, Ph.D.
> >International Health Institute
> >Brown University Box G-8033
> >Providence, RI 02912 USA
> >Jonathan_Kurtis at Brown.edu>>http://www.iol.ie/~users>> Ireland On Line Users' Group
Serology is ideal for INITIAL typing, but in the event of ambiguities
(which are not uncommon with serotyping), it would be best of all to
perform sequence-based typing to SPECIFICALLY clarify what the allele
is. After all, sequence-based typing is the only was of COMPLETELY
clarifying a type, and also of elucidating new allelic forms (many of
which might involve such subtle changes in the floor of the antigen
binding groove that they fail to elicit an ambiguous serological
type...you could therefore COMPLETELY MISS an allelic variant!)...with
B-locus specific primers, you could easily amplify the locus via RT-PCR
of RNA.
Kiley R. Prilliman
University of Oklahoma Health Sciences Center
Oklahoma City
