Entering edit mode
Dear Djie,
I have generalized the removeBatchEffect() function so that it will now accept continuous covariates as well as batch factors. Just enter BSCE as the 'covariates' argument. The updated function will be on the devel version of limma, and I have attached it.
Best wishes
Gordon
> Date: Tue, 31 Jan 2012 13:08:09 +0100
> From: Djie Tjwan Thung <djie.thung at gmail.com>
> To: bioconductor at r-project.org
> Subject: [BioC] Adjusting removeBatchEffect from limma
>
> Dear Gordon Smyth, bioconductor list,
>
> In the limma package we found a function called, removeBatchEffect, which
> removes the effects of batch effects or other technical variables on a gene
> expression matrix. The code of this removeBatchEffect is as follows:
>
> function (x, batch, batch2 = NULL, design = matrix(1, ncol(x), 1)) {
> x <- as.matrix(x)
> batch <- as.factor(batch)
> contrasts(batch) <- contr.sum(levels(batch))
> if (is.null(batch2)) {
> X <- model.matrix(~batch)[, -1, drop = FALSE]
> }
> else {
> batch2 <- as.factor(batch2)
> contrasts(batch2) <- contr.sum(levels(batch2))
> X <- model.matrix(~batch + batch2)[, -1, drop = FALSE]
> }
> X <- qr.resid(qr(design), X)
> t(qr.resid(qr(X), t(x)))
> }
>
> Now we have an ?expression? matrix coming from a methylation array. As
> bisulfite conversion efficiency (BSCE) is a known confounder for this type
> of array, we?d like to adjust the matrix for the confounding effects of
> BSCE. BSCE is a numerical variable, however the removeBatchEffect expects a
> categorical variable as it handles the batch variable as a factor.
>
> We were wondering if its valid to remove the following lines of code in
> this function regarding, batch and batch2, so the function treats these
> variables as numerical variables:
>
> batch <- as.factor(batch)
> contrasts(batch) <- contr.sum(levels(batch))
>
> If someone is willing, you can test out this altered version by loading the
> .RData in the following link:
> http://good.net/dl/au/fa43/contTechnicalVar.RData and using the following
> demo script:
>
> ###
>
> #removeNumericBatchEffect(): altered version of removeBatchEffect(), with the as.factor() and contrasts() lines commented out
>
> #mod: model matrix containing variables of interest (intercept, age and gender)
>
> # head(mod) will give:
>
> # (Intercept) as.factor(pheno$gender)v pheno$age
> #1 1 1 55.9
> #2 1 1 32.0
>
> #create random intensity values for bisulfite conversion
>
> bisulfite.conversion <- sample(6000:10000, 95)
>
> #Run adjusted removeBatchEffect function
>
> removeNumericBatchEffect(expr, batch = bisulfite.conversion, design= mod)
>
> We'd appreciate any input. We?ve also considered ComBat from the sva
> package as an alternative, but as this function also only accepts
> categorical variables, this limma function seems like a better option to
> modify its code for our purpose.
>
> Regards,
>
> Djie Thung
> Bioinformatics intern at UMC Utrecht
> Dept. of Medical Genetics
