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我正在尝试使用 Phyloseq 包的 merge_sample 选项获得相对丰度。

当我用所有样本计算每个 Phylum 的平均值时(我将以 GlobalPatterns 为例);我的意思是,Globalpaters 有 26 个样本,所以我做了类似的东西

library(phyloseq)
library(plyr)
data(GlobalPatterns)
TGroup <- tax_glom(GlobalPatterns, taxrank = "Phylum")
PGroup <- transform_sample_counts(TGroup, function(x)100* x / sum(x))
OTUg <- otu_table(PGroup)
TAXg <- tax_table(PGroup)[,"Phylum"]
AverageD <- as.data.frame(rowMeans(OTUg))
names(AverageD) <- c("Mean")
GTable <- merge(TAXg, AverageD, by=0, all=TRUE)
GTable$Row.names = NULL
GTable <- GTable[order(desc(GTable$Mean)),]
head(GTable)

我得到类似的东西:

        Phylum           Mean

1 Proteobacteria      29.45550
2 Firmicutes          18.87905
3 Bacteroidetes       17.34374
4 Cyanobacteria       13.70639
5 Actinobacteria      8.93446
6....... More.....

我觉得还可以!!!!

但是,当我托盘法师merge_samples(通过:SampleType):

    ps <- tax_glom(GlobalPatterns, "Phylum")
    ps0 <- transform_sample_counts(ps, function(x)100* x / sum(x))
    ps1 <- merge_samples(ps0, "SampleType")
    ps2 <- transform_sample_counts(ps1, function(x)100* x / sum(x))
    ps3 <- ps2
    otu_table(ps3) <- t(otu_table(ps3)) # transpose the matrix otus !!!
    OTUg <- otu_table(ps3)
    TAXg <- tax_table(ps3)[,"Phylum"]
    GTable <- merge(TAXg, OTUg, by=0, all=TRUE)
    GTable$Row.names = NULL
    GTable$Mean=rowMeans(GTable[,-c(1)], na.rm=TRUE)
    GTable <- GTable[order(desc(GTable$Mean)),]
   head(GTable)

我得到相同的税,但在平均值列中具有不同的百分比:

  Phylum Feces Freshwater Freshwater Mock Ocean Sediment Skin Soil Tongue Mean
1 Proteobacteria  1.58 16.71 18.61 20.10 38.00 71.03 31.98 32.66 44.49 30.57
2 Firmicutes 54.82 0.12 0.65 41.42 0.08 2.53 30.67 0.64 21.67 16.96
3 Bacteroidetes 35.23 11.92 5.07 24.97 31.17 7.01 9.09 9.90 12.28 16.29
4 Cyanobacteria 2.63 30.17 62.57 0.16 19.18 3.24 4.65 0.97 6.61 14.46
5 Actinobacteria 3.47 37.11 1.74 8.39 5.12 1.04 16.78 9.99 7.49 10.13

在这一点上,通过 SampleType 的 merge_samples,每一列(样本)都会使分类群变得模糊,并且每个样本中每个门的百分比会发生变化(粪便淡水淡水......),我理解这一点,但每个门的总体平均值必须相同,即使我合并样本,在这种情况下,平均值也不同(Proteobacteria 30.57,Firmicutes 16.9,Bacteroidetes 16.29 ......)。

任何解决方案或建议????

谢谢

4

1 回答 1

2

在第一部分中,您将在所有样本中获取平均值。在第二个中,您正在采用分组均值的平均值。仅当每组的观察次数相同时,这两者才等效。

例如:

# equal n for each group
abundance = seq(0.1,0.6,by=0.1)
group = rep(letters[1:3],each=2)
mean(tapply(abundance,group,mean)) == mean(abundance)
[1] TRUE

# unequal n
abundance = seq(0.1,0.6,by=0.1)
group = rep(letters[1:3],1:3)
mean(tapply(abundance,group,mean)) == mean(abundance)
[1] FALSE

您的每个 SampleType 的 n 不同

TGroup <- tax_glom(GlobalPatterns, taxrank = "Phylum")
PGroup <- transform_sample_counts(TGroup, function(x)100* x / sum(x))
SampleType = sample_data(PGroup)$SampleType
table(SampleType)

SampleType
             Feces         Freshwater Freshwater (creek)               Mock 
                 4                  2                  3                  3 
             Ocean Sediment (estuary)               Skin               Soil 
                 3                  3                  3                  3 
            Tongue 
                 2 

要在样本中获得相同的平均丰度,您需要找到每个 SampleType 的平均丰度,然后求平均值:

mean_PGroup = sapply(levels(SampleType),function(i){
  rowMeans(otu_table(PGroup)[,SampleType==i])
})

phy = tax_table(PGroup)[rownames(mean_PGroup ),"Phylum"]
rownames(mean_PGroup) = phy
head(sort(rowMeans(mean_PGroup),decreasing=TRUE))

 Proteobacteria      Firmicutes   Bacteroidetes   Cyanobacteria  Actinobacteria 
      30.572773       16.956254       16.293286       14.463643       10.126875 
Verrucomicrobia 
       2.774216 
于 2019-11-28T10:37:56.163 回答