Reviewer's report
Title:Differential detection of nuclear envelope autoantibodies in primary biliary
cirrhosis using routine and alternative methods
Version: 1Date:9 January 2010
Reviewer: Marlyn Mayo
Reviewer's report:
Major Compulsory Revisions: none
Minor Essential Revisions: none
Minor Discretionary Revisions:
1. Figure legend 4 does not seem to match labeling of figure (? figure meant to
show purity of nuclear enveloped isolated from rat liver and ? IIF on Hep2
slides?)
2. Please clarify how many of the 103 sera were AMA positive. Just the ones in
Figure 4?
Comment:
1. The question posed by the authors is well defined: How do the different
methods of detecting nuclear envelope Abs in PBC compare with respect to
sensitivity and specificity and what are the reasons for discrepancies between
the different methods?
2. The methodology is sound and thorough. It is described well enough that
others could duplicate their methods.
3. The data confirm several previously reported findings, including an imperfect
correlation between IIF, ELISA and IB for detecting ANEA, the ability of other
autoAbs to interfere with IIF detection of ANEA, that anti-gp210 accounts for
most ANEA. However they also report several distinct findings, including
excellent results with purified rat nuclei IIF and lack of p62 reacitvity. They did
seem to have poorer results with the commerically available ELISA than others
have reported. The article is of interest mainly to pathologists, immunologists, or
hepatologists who utilize these assays. The authors have shown beautiful results
using IIF with isolated nuclei, although this may be too cumbersome a technique
to employ in clinical laboratories. Likewise, the finding that all the AMA positive
sera obscured the ability to see ANEA on IIF, even with their techniques, limits
clinical utility of that assay in PBC patients. Adding a second antigen to the
ELISA and/or using both IgG and IgM secondary antibodies in the IIF assay,
however, are easily done and are excellent recommendations that come from
this work.
4. Data is presented in an unbiased fashion. The number of percentages and
degree of overlap and separation of test results becomes overwhelming at times,
but the data are necessary and the figures help to clarify.
5. The data support the conclusion that current lab techniques can be improved
by adding other substrates.
6. Limitations are not discussed in detail, but there are not many.
7. The introduction refers to the other studies evaluating these assays fairly.
8. The title and abstract clearly convey what has been found.
9. The manuscript is meticulously drafted, albeit lengthy.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declar that I have no competing interests.
Reviewer's report
Title:Differential detection of nuclear envelope autoantibodies in primary biliary
cirrhosis using routine and alternative methods
Version: 1Date:9 January 2010
Reviewer: Marlyn Mayo
Reviewer's report:
Major Compulsory Revisions: none
Minor Essential Revisions: none
Minor Discretionary Revisions:
1. Figure legend 4 does not seem to match labeling of figure (? figure meant to
show purity of nuclear enveloped isolated from rat liver and ? IIF on Hep2
slides?)
2. Please clarify how many of the 103 sera were AMA positive. Just the ones in
Figure 4?
Comment:
1. The question posed by the authors is well defined: How do the different
methods of detecting nuclear envelope Abs in PBC compare with respect to
sensitivity and specificity and what are the reasons for discrepancies between
the different methods?
2. The methodology is sound and thorough. It is described well enough that
others could duplicate their methods.
3. The data confirm several previously reported findings, including an imperfect
correlation between IIF, ELISA and IB for detecting ANEA, the ability of other
autoAbs to interfere with IIF detection of ANEA, that anti-gp210 accounts for
most ANEA. However they also report several distinct findings, including
excellent results with purified rat nuclei IIF and lack of p62 reacitvity. They did
seem to have poorer results with the commerically available ELISA than others
have reported. The article is of interest mainly to pathologists, immunologists, or
hepatologists who utilize these assays. The authors have shown beautiful results
using IIF with isolated nuclei, although this may be too cumbersome a technique
to employ in clinical laboratories. Likewise, the finding that all the AMA positive
sera obscured the ability to see ANEA on IIF, even with their techniques, limits
clinical utility of that assay in PBC patients. Adding a second antigen to the
ELISA and/or using both IgG and IgM secondary antibodies in the IIF assay,
however, are easily done and are excellent recommendations that come from
this work.
4. Data is presented in an unbiased fashion. The number of percentages and
degree of overlap and separation of test results becomes overwhelming at times,
but the data are necessary and the figures help to clarify.
5. The data support the conclusion that current lab techniques can be improved
by adding other substrates.
6. Limitations are not discussed in detail, but there are not many.
7. The introduction refers to the other studies evaluating these assays fairly.
8. The title and abstract clearly convey what has been found.
9. The manuscript is meticulously drafted, albeit lengthy.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declar that I have no competing interests.
Please review this paper.
user
Reviewer's report
Title:Differential detection of nuclear envelope autoantibodies in primary biliary
cirrhosis using routine and alternative methods
Version: 1Date:9 January 2010
Reviewer: Marlyn Mayo
Reviewer's report:
Major Compulsory Revisions: none
Minor Essential Revisions: none
Minor Discretionary Revisions:
1. Figure legend 4 does not seem to match labeling of figure (? figure meant to
show purity of nuclear enveloped isolated from rat liver and ? IIF on Hep2
slides?)
2. Please clarify how many of the 103 sera were AMA positive. Just the ones in
Figure 4?
Comment:
1. The question posed by the authors is well defined: How do the different
methods of detecting nuclear envelope Abs in PBC compare with respect to
sensitivity and specificity and what are the reasons for discrepancies between
the different methods?
2. The methodology is sound and thorough. It is described well enough that
others could duplicate their methods.
3. The data confirm several previously reported findings, including an imperfect
correlation between IIF, ELISA and IB for detecting ANEA, the ability of other
autoAbs to interfere with IIF detection of ANEA, that anti-gp210 accounts for
most ANEA. However they also report several distinct findings, including
excellent results with purified rat nuclei IIF and lack of p62 reacitvity. They did
seem to have poorer results with the commerically available ELISA than others
have reported. The article is of interest mainly to pathologists, immunologists, or
hepatologists who utilize these assays. The authors have shown beautiful results
using IIF with isolated nuclei, although this may be too cumbersome a technique
to employ in clinical laboratories. Likewise, the finding that all the AMA positive
sera obscured the ability to see ANEA on IIF, even with their techniques, limits
clinical utility of that assay in PBC patients. Adding a second antigen to the
ELISA and/or using both IgG and IgM secondary antibodies in the IIF assay,
however, are easily done and are excellent recommendations that come from
this work.
4. Data is presented in an unbiased fashion. The number of percentages and
degree of overlap and separation of test results becomes overwhelming at times,
but the data are necessary and the figures help to clarify.
5. The data support the conclusion that current lab techniques can be improved
by adding other substrates.
6. Limitations are not discussed in detail, but there are not many.
7. The introduction refers to the other studies evaluating these assays fairly.
8. The title and abstract clearly convey what has been found.
9. The manuscript is meticulously drafted, albeit lengthy.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declar that I have no competing interests.
assistant