Name: Nicole Anne F.

Lubis
Year & Section: 2C-MT
SN: 29

Date: April 13, 2015

Subject: Biochemistry Lecture

Journal Article: Validation of a Quantitative Polymerase Chain Reaction Method for

Human

Alu

Gene

Detection

in

Michrochimeric

Pigs

Used

as

Donors

for

Xenotransplantation
Authors: Abellaneda, Martinez-Alarcon, Quereda, et. al.
I. SYNOPSIS
In order to resolve the complex process of xenotransplantation, which was
opposed mainly by immunologic rejection, one of the strategies proposed by
researchers was the generation of chimeric pigs that could promote graft tolerance.
However, it was still a challenge to detect the human cell engraftments in chimeric pigs.
Thus, the study aimed to optimize and to validate the current quantitative polymerase
chain reaction (qPCR) method used in detecting and quantifying human specific DNA
elements called Alu gene in tissues and blood samples of pigs in which human stem
cells were engrafted. A mixture of human DNA and different quantities of porcine DNA
was subjected to qPCR for quantification. By generating a reproducible and bimodal
melting curve, qPCR was proven to be a specific and strong technique in detecting
human cell engraftments in piglets after human cell injection.
II. CRITICAL REVIEW
The article provided a clear and comprehensible method, result and discussion in
the validation of the efficiency of quantitative polymerase chain reaction (qPCR) in
detecting human cell engraftments in piglets following human cell injection. The
breakdown of facts, theories, steps and processes was well-structured and clearly
presented. The article was preceded by affirming the importance of xenotransplantation
in adhering to the problem of organ shortage for transplantation. The previously said
technique was supported by ideas like the generation of chimeric pigs in promoting graft
tolerance and the use of qPCR for identification and quantification of Alu genes in pigs
that helped the readers to easily understand the essentiality of testing the competence
of qPCR.
Some of the strengths of this article were: one, it defined terms and principles
that may trigger confusion to the readers, thus eliminating vague cohesion among

statements; two, it relayed the complete quantitative results for DNA mixes by
formulating a table, and three, the discussion of results flowed smoothly and was
administered by comparing and contrasting various test samples which enabled the
readers to easily sort out information and to arrive to the best conclusion.
On the other hand, there were certain weak points that were present in the
article. Firstly, the data for the regression formula in correcting the subquantification of
high quantities of porcine DNA was not shown. It might have been better to show it in a
tabulated form in order for the readers to see and censure the validity of the used
regression formula. Lastly, the dissociation curve in the assay that produced a bimodal
melting curve was also not shown. The said figure might have been very useful for the
readers in deciphering the relationship among the test samples that were subjected in
temperature change.
Overall, the journal article was well-organized and provided compact evidence in
validating qPCR as a powerful tool to detect and quantify human cell engraftments in
piglets after human stem cell injection.

Journal Article Source: http://0-www.sciencedirect.com.ustlib.ust.edu.ph/science/article/pii/S0041134514012688