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global analysis_COGUK.R
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global analysis_COGUK.R
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# GLOBAL ANALYSIS OF SARS-Cov2 VARIANTS OF CONCERN & INTEREST USING MULTINOMIAL SPLINE FITS ####
# DATA: COG-UK, NextCladePangolineages called by DaveMc Nally, see https://ukcovid.xyz/variant-landscape.php and https://ukcovid.xyz/cog.csv ####
# T. Wenseleers
# last update 9 FEBRUARY 2023
# for similar analysis see https://nbviewer.org/github/gerstung-lab/SARS-CoV-2-International/blob/main/genomicsurveillance-int.ipynb#Check-some-fast-growing-lineages
# rm(list = ls()) # clear workspace
gc()
# set data source: open NCBI data or GISAID data ####
source = "COGUK" # or "GISAID" (CovSpectrum) or "NCBI", using COG-UK data here
# set COVSPECTRUM credentials if you would like to use private GISAID data ####
# do this by setting REACT_APP_LAPIS_ACCESS_KEY as an environment variable
# using Sys.setenv(REACT_APP_LAPIS_ACCESS_KEY = "XXXX")
# cf https://cov-spectrum.org/static/js/main.b4ab11d7.js
# (using open NCBI data is also possible by setting source="NCBI" and then no key is needed)
if (file.exists("..//set_COVSPECTRUM_credentials.R")) source("..//set_COVSPECTRUM_credentials.R")
# load (& if needed install) required packages & load some utility function ###
# install.packages("pacman")
library(pacman)
pacman::p_load(devtools, nnet, splines, pracma, readr, ggplot2, ggthemes, scales,
archive, dplyr, stringr, lubridate, tidyr, countrycode,
memoise, readxl, covidregionaldata, tidyquant, data.table, R.utils,
locatexec, pals, inspectdf, zoo, RSelenium, jsonlite)
Sys.setenv(GITHUB_PAT = "")
if (!require(marginaleffects)) devtools::install_github("tomwenseleers/marginaleffects")
require(marginaleffects)
pacman::p_load_gh("melff/mclogit/pkg", "rvlenth/emmeans",
"epiforecasts/covidregionaldata",
"melff/mclogit/pkg")
# load some utility functions to get aggregated NextcladePangolin lineage frequencies from covSPECTRUM
source(".//download_covSpectrum.R") # utility function download_covSpectrum to download data from covSpectrum
# select target variant (CovSpectrum nextcladePangoLineage notation) ####
target_variant = "XBB.1.5*"
# select countries to use in analyses, here those with >=5 XBB.1.5* sequences ####
# over the past 30 days & 100 sequenced genomes in total over that period
minseqs = 5
mintotalseqs = 100
lastdays = 30
if (source=="COGUK") { countries = "United Kingdom" } else {
toplist = countrieswithvariant(target_variant="XBB.1.5*",
minseqs=minseqs,
mintotalseqs=mintotalseqs,
lastdays=lastdays)
toplist
countries = unique(toplist$country)
}
# define countries for which we would like to divide by division (state/province) ####
countries_bydivision = c("USA", "United Kingdom") # could be good for India too, but names need some fixing
# download variant data ####
date_from = "2019-12-01"
if (source!="COGUK") { # CovSpectrum GISAID data
data_percountry = bind_rows(lapply(countries[!countries %in% countries_bydivision],
function (country) download_covSpectrum(source=source,
date_from=date_from,
country=country, # vector of country names, NA for all
nextcladePangoLineages=NA, # get all lineages
bydate=TRUE,
bypangolineage=TRUE,
bydivision=FALSE)
))
data_percountry$division = data_percountry$country
data_percountry_perdivision = bind_rows(lapply(countries[countries %in% countries_bydivision],
function (country) download_covSpectrum(source=source,
date_from=date_from,
country=country, # vector of country names, NA for all
nextcladePangoLineages=NA, # get all lineages
bydate=TRUE,
bypangolineage=TRUE,
bydivision=TRUE)
))
sort(unique(data_percountry_perdivision$division))
data_percountry_perdivision$division = gsub("Stockton", "California", data_percountry_perdivision$division, fixed=T)
data_percountry_perdivision$division = gsub("Washington DC", "Washington", data_percountry_perdivision$division, fixed=T)
data_percountry_perdivision = data_percountry_perdivision[!data_percountry_perdivision$division %in%
c(NA, "Un", "un", "USA", "Stockton", "American Samoa",
"Guam", "Northern Mariana Islands",
"Puerto Rico", "Virgin Islands",
"Anguilla","Birmingham",
"British Virgin Islands",
"Cayman Islands",
"Gibraltar",
"Montserrat",
"Turks and Caicos Islands"),]
} else { # COG-UK data
data_percountry_perdivision = read.csv("https://ukcovid.xyz/cog.csv", header=F) %>%
rename(id = V1,
date = V2,
nextcladePangoLineage = V3
)
subsettoONS = FALSE # subset to ONS samples?
if (subsettoONS) data_percountry_perdivision = data_percountry_perdivision[grepl("QEUH", data_percountry_perdivision$id),]
data_percountry_perdivision$division = str_extract(data_percountry_perdivision$id, "^[^/]+")
data_percountry_perdivision$region = "United Kingdom"
data_percountry_perdivision$count = 1
data = data_percountry_perdivision
}
if (source!="COGUK") {
# aggregate US states into Northeast, South, West, Midwest ?
aggregate_divisions = TRUE
keyval= read.csv("./data/division_aggregation.csv")
sort(unique(data_percountry_perdivision$division))[!sort(unique(data_percountry_perdivision$division)) %in% keyval$division] # OK
if (aggregate_divisions) {
data_percountry_perdivision = data_percountry_perdivision %>%
left_join(keyval) %>%
mutate(division = value) %>%
select(-value)
}
data = bind_rows(data_percountry, data_percountry_perdivision)
}
# convert date to date format ####
data$date = as.Date(fast_strptime(data$date, "%Y-%m-%d")) # faster than as.Date(data$date)
# define output directories & info to put on plots if desired ####
today = as.Date(Sys.time())
today_num = as.numeric(today)
if (source!="COGUK") target_dir = "./data/GISAID" else target_dir = "./data/COGUK"
suppressWarnings(dir.create(target_dir))
if (source!="COGUK") plotdir = "./plots/GISAID" else plotdir = "./plots/COGUK"
suppressWarnings(dir.create(plotdir))
tag = paste("@TWenseleers\n",today)
# define main variant lineages ####
# I need to code this in a more generic way as in
# https://nbviewer.org/github/gerstung-lab/SARS-CoV-2-International/blob/main/genomicsurveillance-int.ipynb#Check-some-fast-growing-lineages
# lineage_aliases = fromJSON("https://cov-spectrum.org/api/v2/resource/pango-lineage-alias")
# some regular expression helper functions
# lineage
lin = function (lineage, x=data$nextcladePangoLineage) { pat = paste0("^", gsub(".","\\.",lineage, fixed=T), "$")
grepl(pat, x, fixed=F, perl=T) }
# lineage plus sublineages
linplus = function (lineage, x=data$nextcladePangoLineage) { pat = paste0("^", gsub(".","\\.",lineage, fixed=T), "$","|",paste0("^", gsub(".","\\.",lineage, fixed=T)))
grepl(pat, x, fixed=F, perl=T) }
# one of X lineages
lin_oneof = function (lineages, x=data$nextcladePangoLineage) {
rowSums(sapply(lineages, function (lineage) {
pat = paste0("^", gsub(".","\\.",lineage, fixed=T), "$")
grepl(pat, x, fixed=F, perl=T) }))>=1
}
# e.g. lin_oneof(c("BA.5","BF.7"))
# one of X lineages plus sublineages
linplus_oneof = function (lineages, x=data$nextcladePangoLineage) {
rowSums(sapply(lineages, function (lineage) {
pat = paste0("^", gsub(".","\\.",lineage, fixed=T), "$","|",paste0("^", gsub(".","\\.",lineage, fixed=T)))
grepl(pat, x, fixed=F, perl=T) }))>=1
}
# linplus_oneof(c("BA.5","BF.7"))
# date from XX
datefrom = function (date, x=data$date) { x >= as.Date(date) }
# datefrom(c("2021-01-01"))
# define lineages
levels_VARIANTS = c("Other", "B.1.177 (EU1)", "B.1.160 (EU2)",
"B.1.221 (20A/S:98F)", "Beta", "Alpha", "Delta",
"Omicron (BA.1*)", "Omicron (BA.2*)",
"Omicron (BA.2.12.1*)",
"Omicron (BA.2.38*)",
"Omicron (BA.4*)", "Omicron (BA.4.6*)",
"Omicron (BA.5*)", "Omicron (BA.5.2*)", "Omicron (BF.7*)",
"Omicron (BA.2.76*)",
"Omicron (BA.2.75*)",
"Omicron (BQ.1.1*)",
"Omicron (BQ.1*)",
"Omicron (BR.2.1*)",
"Omicron (CH.1.1*)",
"Deltacron (XAY.2*)",
"Omicron (XBF*)",
# "Omicron (XBK*)", # TO DO XBK* = BM.1.1* (Nextclade) + C1627T + S:F486P = https://cov-spectrum.org/explore/World/AllSamples/Past6M/variants?aaMutations=S%3AF486P&nucMutations=C1627T&nextcladePangoLineage=BM.1.1*
"Omicron (XBB*)",
"Omicron (XBB.1.5*)",
"Omicron (XBB.1.9.1*)"
)
target_variant = "Omicron (XBB.1.5*)"
baseline = "Omicron (BQ.1.1*)"
# I am using this order in plots, baseline is in fits coded as reference level
data$variant <- case_when(
linplus("XBB.1.9") ~ "Omicron (XBB.1.9*)",
linplus("XBB.1.5") ~ "Omicron (XBB.1.5*)",
linplus("XBB") ~ "Omicron (XBB*)",
linplus("BR.2.1") ~ "Omicron (BR.2.1*)",
linplus("XBF") ~ "Omicron (XBF*)",
linplus("CH.1.1") ~ "Omicron (CH.1.1*)",
linplus("XAY.2") ~ "Deltacron (XAY.2*)",
linplus("BQ.1.1")&
datefrom("2022-02-01") ~ "Omicron (BQ.1.1*)",
linplus("BQ.1")&
datefrom("2022-02-01") ~ "Omicron (BQ.1*)",
linplus("BA.2.75")&
(!(linplus("BN")|linplus("BM")|linplus("BR")|linplus("BL")|linplus("CH")|linplus("BQ")))&
datefrom("2022-04-01") ~ "Omicron (BA.2.75*)",
linplus("BA.2.76") ~ "Omicron (BA.2.76*)",
linplus("BF.7")&
datefrom("2022-02-01") ~ "Omicron (BF.7*)",
linplus("BA.5.2")&
datefrom("2022-02-01") ~ "Omicron (BA.5.2*)",
linplus("BA.5")&
datefrom("2021-12-01") ~ "Omicron (BA.5*)",
linplus("BA.4.6")&
datefrom("2021-12-01") ~ "Omicron (BA.4.6*)",
linplus("BA.4")&
datefrom("2021-12-01") ~ "Omicron (BA.4*)",
linplus("BA.2.12.1")&
datefrom("2021-12-01") ~ "Omicron (BA.2.12.1*)",
linplus("BA.2.38")&
datefrom("2021-09-01") ~ "Omicron (BA.2.38*)",
linplus("BA.2")&
datefrom("2021-09-01") ~ "Omicron (BA.2*)",
linplus("BA.1")&
datefrom("2021-09-01") ~ "Omicron (BA.1*)",
linplus("B.1.617.2")|linplus("AY")&
datefrom("2020-10-30") ~ "Delta",
linplus("B.1.1.7")&
datefrom("2020-09-20") ~ "Alpha",
linplus("B.1.351")&
datefrom("2020-08-10") ~ "Beta",
linplus("B.1.221")&
datefrom("2020-03-01") ~ "B.1.221 (20A/S:98F)",
linplus("B.1.160")&
datefrom("2020-02-15") ~ "B.1.160 (EU2)",
linplus("B.1.177")&
datefrom("2020-05-27") ~ "B.1.177 (EU1)",
TRUE ~ "Other"
)
length(unique(data$variant)) == length(levels_VARIANTS) # correct
max(data$date) # "2023-01-13"
# note: in India BA.2.38 & BA.2.38.1 caused an infection wave in some states - hence separated out above
# B.1.177+B.1.160+B.1.221 were behind the 2020 wave in fall in Europe & each had one spike mutations & a small growth rate advantage relative to predominant B.1.1
# earliest realistic dates were taken from
# https://raw.githubusercontent.com/nextstrain/ncov/master/defaults/clade_emergence_dates.tsv
# define lineage colours
n = length(levels_VARIANTS)
lineage_cols = case_when(
levels_VARIANTS=="Other" ~ "grey65",
levels_VARIANTS=="B.1.177 (EU1)" ~ "darkorange4",
levels_VARIANTS=="B.1.160 (EU2)" ~ "darkorange3",
levels_VARIANTS=="B.1.221 (20A/S:98F)" ~ "darkorange2",
levels_VARIANTS=="Beta" ~ "green4",
levels_VARIANTS=="Alpha" ~ "#0085FF",
levels_VARIANTS=="Delta" ~ "mediumorchid",
levels_VARIANTS=="Omicron (BA.1*)" ~ "red",
levels_VARIANTS=="Omicron (BA.2*)" ~ "red3",
levels_VARIANTS=="Omicron (BA.2.12.1*)" ~ "black",
levels_VARIANTS=="Omicron (BA.2.38*)" ~ "red4",
levels_VARIANTS=="Omicron (BA.4*)" ~ "green3",
levels_VARIANTS=="Omicron (BA.4.6*)" ~ "green2",
levels_VARIANTS=="Omicron (BA.5*)" ~ "blue4",
levels_VARIANTS=="Omicron (BA.5.2*)" ~ "blue3",
levels_VARIANTS=="Omicron (BF.7*)" ~ "dodgerblue",
levels_VARIANTS=="Omicron (BA.2.76*)" ~ "magenta4",
levels_VARIANTS=="Omicron (BA.2.75*)" ~ "magenta",
levels_VARIANTS=="Omicron (BQ.1*)" ~ "orange",
levels_VARIANTS=="Omicron (BQ.1.1*)" ~ "orange2",
levels_VARIANTS=="Omicron (BR.2.1*)" ~ "orange3",
levels_VARIANTS=="Omicron (CH.1.1*)" ~ "yellow3",
levels_VARIANTS=="Deltacron (XAY.2*)" ~ "cyan4",
levels_VARIANTS=="Omicron (XBF*)" ~ "cyan3",
levels_VARIANTS=="Omicron (XBB*)" ~ "cyan2",
levels_VARIANTS=="Omicron (XBB.1.5*)" ~ "cyan",
levels_VARIANTS=="Omicron (XBB.1.9.1*)" ~ "yellow"
)
names(lineage_cols) = levels_VARIANTS
length(levels_VARIANTS) == length(lineage_cols) # correct
pal.bands(lineage_cols)
data$variant = factor(data$variant, levels=levels_VARIANTS)
table(data$variant)
table(data$region, data$variant)
table(data$division, data$variant)
# add week, year & start of week ####
# TO DO: add these columns in download functions?
data$Week = lubridate::week(data$date)
data$Year = lubridate::year(data$date)
data$Year_Week = interaction(data$Year,data$Week)
data$floor_date = fast_strptime(as.character(cut(data$date, "week")), "%Y-%m-%d") # start of week
data$date_num = as.numeric(data$date)
# sort countries by incidence of target variant
# data$country = factor(data$country, levels=toplist$country)
# sort divisions/states & countries by incidence of target variant
total_count_target_variant = data %>%
filter(date>=(as.Date(Sys.time())-lastdays)) %>%
group_by(variant, division) %>%
summarise(total_count_target_variant = sum(count)) %>%
filter(variant==target_variant)
total_n = data %>%
filter(date>=(as.Date(Sys.time())-lastdays)) %>%
group_by(division) %>%
summarise(total_n = sum(count))
total_count_target_variant = left_join(total_count_target_variant, total_n) %>%
mutate(prop_target_variant = total_count_target_variant / total_n) %>%
arrange(desc(prop_target_variant))
data$division = factor(data$division, levels=c(total_count_target_variant$division,
sort(unique(data$division[!data$division %in% total_count_target_variant$division]))))
table(data$division, data$variant)
if (source!="COGUK") { levels_regions = c("Asia","North America","Europe","Africa","South America","Oceania") } else {
levels_regions = c("United Kingdom") }
data$region = factor(data$region, levels=levels_regions)
data$region = droplevels(data$region)
table(data$region, data$variant)
table(data$division, data$variant)
# 2. COMBINED GLOBAL ANALYSIS USING MULTINOMIAL SPLINE FITS ####
# AGGREGATED DATA BY WEEK ####
data_agbyweek1 = data %>%
group_by(variant, floor_date) %>%
summarise(count = sum(count))
data_agbyweek1_total = data %>%
group_by(floor_date) %>%
summarise(total = sum(count))
data_agbyweek1 = left_join(data_agbyweek1, data_agbyweek1_total) %>%
rename(date = floor_date) %>%
mutate(prop = count/total)
data_agbyweek1 = as.data.frame(data_agbyweek1)
data_agbyweek1$date = as.Date(data_agbyweek1$date)
data_agbyweek1$date_num = as.numeric(data_agbyweek1$date)
# write.csv(data_agbyweek1, file=".//data//COVSPECTRUM//COVSPECTRUM aggregated counts by week_all.csv", row.names=F)
# AGGREGATED DATA BY WEEK & COUNTRY/DIVISION ####
data_agbyweekcountry1 = data %>%
group_by(division, variant, floor_date) %>%
summarise(count = sum(count))
data_agbyweekcountry1_total = data %>%
group_by(division, floor_date) %>%
summarise(total = sum(count))
data_agbyweekcountry1 = left_join(data_agbyweekcountry1, data_agbyweekcountry1_total) %>%
rename(date = floor_date) %>%
mutate(prop = count/total)
data_agbyweekcountry1 = as.data.frame(data_agbyweekcountry1)
data_agbyweekcountry1$date = as.Date(data_agbyweekcountry1$date)
data_agbyweekcountry1$date_num = as.numeric(data_agbyweekcountry1$date)
data_agbyweekcountry1$region = data$region[match(data_agbyweekcountry1$division, data$division)]
# write.csv(data_agbyweekcountry1, file="./data/COVSPECTRUM/COVSPECTRUM aggregated counts by start of week and lineage_all.csv", row.names=F)
gc()
# MULLER PLOT (RAW DATA, selected countries pooled, but with big sampling biases across countries/divisions) ####
data_agbyweek1$variant = factor(data_agbyweek1$variant, levels=levels_VARIANTS)
if (source=="COGUK") { brks = "1 week"
lbls = "%d %b %Y" } else {
brks = "1 month"
lbls = "%b %Y" }
muller_raw_all = ggplot(data=data_agbyweek1, aes(x=date, y=count, group=variant)) +
geom_area(aes(lwd=I(1.2), colour=NULL, fill=variant, group=variant), position="fill") +
scale_fill_manual("", values=lineage_cols) +
scale_x_date(date_breaks = brks, date_labels = lbls) +
theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
# guides(color = guide_legend(reverse=F, nrow=2, byrow=T), fill = guide_legend(reverse=F, nrow=2, byrow=T)) +
theme_hc() +
ylab("Share") +
theme(legend.position="right",
axis.title.x=element_blank()) +
labs(title = "SARS-CoV2 LINEAGE FREQUENCIES",
subtitle=paste0("Raw GISAID data up to ",today," (using NextClade Pangolin lineages)")) +
coord_cartesian(xlim=c(min(data$date),max(data$date)), expand=c(0)) +
labs(tag = tag) + theme(plot.tag.position = "bottomright", plot.tag = element_text(vjust = 1, hjust = 1, size=8))
muller_raw_all
ggsave(file=file.path(plotdir,"muller plot_raw data.png"), width=12, height=5)
# FIT NNET::MULTINOM MULTINOMIAL SPLINE MODEL ####
# code uses baseline lineage as reference level
data_agbyweekcountry1$variant = relevel(data_agbyweekcountry1$variant, ref=baseline)
set.seed(1)
gc()
if (source!="COGUK") {
system.time(fit <- nnet::multinom(variant ~ ns(date_num, df=2)+ns(date_num, df=2):region+division,
weights=count,
data=data_agbyweekcountry1,
maxit=10000, MaxNWts=100000)) # iter 1530, 134s
# syntax of model to put on plot legend
model = "variant ~ ns(date, df=2)+ns(date, df=2):region+division"
} else {
# system.time(fit <- nnet::multinom(variant ~ ns(date_num, df=2)*division,
# weights=count,
# data=data_agbyweekcountry1,
# maxit=10000, MaxNWts=100000)) # iter 200, 0.09s
# # syntax of model to put on plot legend
# model = "variant ~ ns(date, df=2)*country"
data_agbyweekcountry1$variant = droplevels(data_agbyweekcountry1$variant)
system.time(fit <- nnet::multinom(variant ~ date_num+division,
weights=count,
data=data_agbyweekcountry1,
maxit=10000, MaxNWts=100000)) # iter 200, 0.09s
model = "variant ~ date+country"
}
# TO DO: change to mclogit::mblogit fit (can take into account overdispersion &
# latest github version now runs OK) or
# the MGLM package (MGLMreg or with regularisation MGLMsparsereg, both allow for overdispersion with
# dist="DM" or dist="NegMN") - but that one gave fitting errors
# # NOTE
# # mblogit & MGLM syntax shown on small data subset
# datsubs = data_agbyweekcountry1[(data_agbyweekcountry1$variant %in% c("Omicron (BQ.1*)",
# "Omicron (XBB.1.5*)",
# "Omicron (XBB*)")) &
# data_agbyweekcountry1$date>=as.Date("2022-08-01"),]
# datsubs$variant = droplevels(datsubs$variant)
# datsubs$region = droplevels(datsubs$region)
# datsubs$division = droplevels(datsubs$division)
# saveRDS(datsubs, file="../data.rds")
# datsubs = readRDS(file="../data.rds")
#
# # example dataset
# download.file("https://www.dropbox.com/s/o6iu51wu7x90omd/data.rds?dl=1",
# "../data.rds",
# method = "auto", mode="wb")
# datsubs = readRDS(file = "../data.rds")
#
# # first nnet::multinom : runs OK
# library(nnet)
# fit_nnet <- nnet::multinom(variant ~ ns(date_num, df=2) + ns(date_num, df=2):region + division,
# weights = count,
# data = datsubs,
# maxit = 10000, MaxNWts = 100000)
# # hist(coef(fit_nnet))
# # range(coef(fit_nnet))
#
# # syntax for mblogit multinomial fit with overdispersion is
# devtools::install_github("melff/mclogit",subdir="pkg")
# library(mclogit)
# fit_mblogit <- mblogit(formula=variant ~ ns(date_num, df=2) + ns(date_num, df=2):region + division, # runs OK too
# weights=count,
# data=datsubs,
# from.table=TRUE,
# dispersion=TRUE, # fit overdispersion?
# control=mclogit.control(maxit=10000))
# # hist(coef(fit_mblogit))
# # range(coef(fit_mblogit))
#
# # syntax for MGLM should be the following (I think), but returns an error
# library(MGLM)
# datsubs_wide <- datsubs %>%
# pivot_wider(names_from = variant,
# values_from = count) %>%
# replace(is.na(.), 0)
# Y <- as.matrix(datsubs_wide %>% select(tail(names(.), length(unique(datsubs$variant)) )))
# Y <- sweep(Y, 1, FUN="/", rowSums(Y))
# X <- model.matrix(formula(~ns(date_num, df=2) + ns(date_num, df=2):region + division), data=datsubs_wide)
# fit_MGLM <- MGLM::MGLMreg(Y ~ X,
# dist = "MN", # or DMN for Dirichlet Multinomial or NegMN for Negative Multinomial, see also MGLMsparsereg
# weight = datsubs_wide$count,
# maxiters = 10000)
# # gives error
# # Error in if (is.nan(ll.Newton) || ll.Newton >= 0) { :
# # missing value where TRUE/FALSE needed
# # In addition: There were 41 warnings (use warnings() to see them)
# model to use below - I just fitted 1 possible model now - one could vary df etc
fit_best = fit
# we calculate the Hessian using my own faster Rcpp Kronecker-product based function
source(".//fastmultinomHess.R") # faster way to calculation Hessian of multinomial fits
gc()
system.time(fit_best$Hessian <- fastmultinomHess(fit_best, model.matrix(fit_best))) # 7s
# we add variance-covariance matrix as extra slot to be re-used later
system.time(fit_best$vcov <- vcov(fit_best)) # 1s
gc()
# save environment
# save.image("./environment_COGUK.RData")
# load("./environment_COGUK.RData")
# save multinom fit
if (source!="COGUK") saveRDS(fit_best, file="./fits/multinom_fit.rds") else saveRDS(fit_best, file="./fits/multinom_fit_COGUK.rds")
# CALCULATE GROWTH RATE ADVANTAGE OVER BASELINE REFERENCE LEVEL BQ.1.1* TODAY ####
# with new faster marginaleffects code
library(marginaleffects)
system.time(meffects <- marginaleffects(fit_best,
type = "link", # = additive log-ratio = growth rate advantage relative to BQ.1*
variables = c("date_num"),
by = c("group"),
vcov = fit_best$vcov,
newdata = datagrid(date_num = today_num
))) # 15s
# growth rate advantage compared to reference level by region
if (source != "COGUK") {
system.time(meffects_byregion <- marginaleffects(fit_best,
type = "link", # = additive log-ratio = growth rate advantage relative to BQ.1*
variables = c("date_num"),
by = c("group", "region"),
vcov = fit_best$vcov,
newdata = datagrid(date_num = today_num,
region = unique(data_agbyweekcountry1$region)
))) # 15s
} else {
system.time(meffects_byregion <- marginaleffects(fit_best,
type = "link", # = additive log-ratio = growth rate advantage relative to BQ.1*
variables = c("date_num"),
by = c("group", "division"),
vcov = fit_best$vcov,
newdata = datagrid(date_num = today_num,
division = unique(data_agbyweekcountry1$division)
))) # 15s
meffects_byregion = meffects_byregion %>%
rename(region = division)
}
# for all pairwise growth rate differences:
# growth_differences = comparisons(
# fit_best,
# newdata = datagrid(date_num = today_num),
# variables = "date_num",
# by = "region",
# type = "clr", # here we could either use "clr" (centered logratio) or "link" (additive logratio) - this gives same result
# hypothesis = "pairwise")
# old emtrends code to calculate pairwise growth rate differences
# system.time(emtr_pairw <- emtrends(fit_best, revpairwise ~ variant,
# by="region",
# var="date_num", mode="latent",
# at=list(date_num=today_num))) #
# delta_r_pairw = data.frame(confint(emtr_pairw,
# adjust="none", df=NA)$contrasts,
# p.value=as.data.frame(emtr_pairw$contrasts)$p.value)
# delta_r_pairw
# write.csv(delta_r_pairw, file.path(plotdir, "growth rate advantage all variants vs BA_5_2.csv"), row.names=F)
# plot of growth rate advantage of last n newest variants
# TO DO: order by selective advantage and then take top lastn
lastn = 14 # last n variants to show - change in top n ?
sel_variants = tail(levels_VARIANTS,lastn)
sel_variants = sel_variants[!sel_variants %in% c(baseline, "Omicron (BA.4.6*)", "Omicron (BA.2.76*)")]
meffects_sel1 = meffects[meffects$group %in% sel_variants,]
meffects_sel1$group = factor(meffects_sel1$group, levels=meffects_sel1$group[order(meffects_sel1$dydx, decreasing=T)])
cols = colorRampPalette(c("red3", "blue3"))(length(levels(meffects_sel1$group)))
if (source!="COGUK") { subtit = paste0("based on multinomial fit ", model, "\nGISAID data with NextcladePangolin lineage definition,\nusing data from countries with >=", minseqs, " XBB.1.5* sequences") } else {
subtit = paste0("based on multinomial fit ", model, "\nCOG-UK data with NextcladePangolin lineages called by Dave McNally")
if (subsettoONS) subtit = paste0(subtit, " (ONS subset)")
}
meffects_sel1
write.csv(meffects_sel1, file.path(plotdir, "growth rate advantage all variants vs BQ_1_1.csv"), row.names=F)
qplot(data=meffects_sel1,
x=group, y=dydx*100, ymin=conf.low*100, ymax=conf.high*100, fill=group, geom="col",
width=I(0.7)) +
geom_linerange(aes(lwd=I(0.4))) + ylab("Growth rate advantage\nrelative to BQ.1.1* (% per day)") +
scale_fill_manual(values=cols) +
theme(legend.position="none") + xlab("") +
ggtitle("GROWTH RATE ADVANTAGE OF SARS-CoV2 VARIANTS",
subtitle=subtit ) +
labs(tag = tag) +
theme(plot.tag.position = "bottomright",
plot.tag = element_text(vjust = 1, hjust = 1, size=8)) +
theme(axis.text.x = element_text(angle = 45, hjust=1))
if (!subsettoONS) ggsave(file=file.path(plotdir,"growth rate advantage VOCs_overall.png"), width=7, height=5) else ggsave(file=file.path(plotdir,"growth rate advantage VOCs_overall_ONS.png"), width=7, height=5)
# plot of growth rate advantage of last X newest variants by continent
# TO DO: order by selective advantage and then take top n
sel_variants = tail(levels_VARIANTS,lastn)
sel_variants = sel_variants[!sel_variants %in% c(baseline, "Omicron (BA.4.6*)", "Omicron (BA.5*)", "Omicron (BA.2.76*)")]
sel_regions = unique(meffects_byregion$region)
# sel_regions = sel_regions[!sel_regions %in% c("Africa")] # too little data
meffects_sel2 = meffects_byregion[meffects_byregion$region %in% sel_regions,]
meffects_sel2 = meffects_sel2[meffects_sel2$group %in% sel_variants,]
meffects_sel2$group = factor(meffects_sel2$group, levels=levels(meffects_sel1$group))
# outlier = (abs(meffects_sel2$dydx)>=0.35)|(meffects_sel2$dydx<0) # typically due to there being too little data
# meffects_sel2 = meffects_sel2[!outlier,]
if (source=="COGUK") div="division" else div="region"
tbl = as.data.frame(table(data[data$variant %in% sel_variants,div],
data[data$variant %in% sel_variants,"variant"]))
colnames(tbl) = c("region", "variant", "count")
meffects_sel2$count = tbl$count[match(interaction(meffects_sel2$region, meffects_sel2$group),
interaction(tbl$region, tbl$variant))]
if (source!="COGUK") reg = "Europe" else reg = "England" # sort by growth advantages in this region
meffects_sel2$group = factor(meffects_sel2$group,
levels=meffects_sel2[meffects_sel2$region==reg,"group"][order(meffects_sel2[meffects_sel2$region==reg,"dydx"],
decreasing=TRUE)])
# retain only estimates with total count > minvariantseqs per region/continet
minvariantseqs = 30
meffects_sel2 = meffects_sel2[meffects_sel2$count>=minvariantseqs, ]
nregions = length(unique(meffects_sel2$region))
qplot(data=meffects_sel2,
x=group, y=dydx*100, ymin=conf.low*100, ymax=conf.high*100, fill=group, geom="col",
width=I(0.7)) +
facet_wrap(~ region, ncol=1) +
geom_linerange(aes(lwd=I(0.4))) + ylab("Growth rate advantage\nrelative to BQ.1* (% per day)") +
scale_fill_manual(values=cols) +
theme(legend.position="none") + xlab("") +
ggtitle("GROWTH RATE ADVANTAGE OF SARS-CoV2 VARIANTS",
subtitle=subtit ) +
labs(tag = tag) +
theme(plot.tag.position = "bottomright",
plot.tag = element_text(vjust = 1, hjust = 1, size=8)) +
theme(axis.text.x = element_text(angle = 45, hjust=1))
ggsave(file=file.path(plotdir,"growth rate advantage VOCs_by region.png"), width=7, height=2*nregions)
# PLOT MULTINOMIAL FIT ####
extrapolate = 14
# date.from = as.numeric(as.Date("2020-01-01"))
date.from = as.numeric(min(data$date)) # as.numeric(as.Date("2021-01-01"))
date.to = today_num+extrapolate
# multinomial model predictions by country/divisions with CIs calculated using margineffects::predictions
step=1
predgrid = expand.grid(list(date_num=as.numeric(seq(date.from, date.to, by=step)),
division=unique(data_agbyweekcountry1$division)))
predgrid$region = data_agbyweekcountry1$region[match(predgrid$division,
data_agbyweekcountry1$division)]
# note: now using Delta method on response scale, better would be to
# calculate CIs as in Effects package on link scale (type="link") or
# on even better on isometric logratio scale (type="ilr") & then backtransform
# but still having some problems with over/underflows with
# type="link" and type="ilr"=isometric logratio is a bit more hassle to backtransform
# (though working correctly)
# rm(fit_preds)
gc()
system.time(fit_preds <- data.frame(predictions(fit_best,
newdata = predgrid,
type = "probs",
vcov = fit_best$vcov))) # %>% # 178s
# transform(conf.low = predicted - 1.96 * std.error,
# conf.high = predicted + 1.96 * std.error) %>%
# group_by(rowid) |>
#mutate_at(c("predicted", "conf.low", "conf.high"), function (x) plogis(x)))
range(fit_preds$predicted)
gc()
fit_preds$conf.high[fit_preds$conf.high>0.99999] = 0.99999 # slight artefact of Delta method on response scale
# fit_preds$conf.high[fit_preds$conf.high<1E-10] = 1E-10
fit_preds$conf.low[fit_preds$conf.low<1E-10] = 1E-10
# fit_preds$conf.low[fit_preds$conf.low>0.99999-10] = 0.99999
# fit_preds$predicted[fit_preds$predicted>0.99999] = 0.99999
# fit_preds$predicted[fit_preds$predicted<1E-10] = 1E-10
# replace NAs by 0
# fit_preds <- fit_preds %>% mutate(predicted = ifelse(is.na(predicted), 0, predicted),
# conf.low = ifelse(is.na(conf.low), 0, conf.low),
# conf.high = ifelse(is.na(conf.high), 0, conf.high))
fit_preds$date = as.Date(fit_preds$date_num, origin="1970-01-01")
fit_preds$variant = NULL
fit_preds = fit_preds %>% rename(variant = group)
fit_preds$variant = factor(fit_preds$variant, levels=levels_VARIANTS)
# TO DO: fix bug with type="link" where some predictions come out as NA,
# and/or switch to type="ilr" - check in my marginaleffects fork
if (source!="COGUK") {
write_csv(fit_preds, file=file.path(plotdir, "COVSPECTRUM fitted lineage frequencies global multinomial fit.csv"))
} else {
write_csv(fit_preds, file=file.path(plotdir, "COGUK fitted lineage frequencies global multinomial fit.csv"))
}
# saveRDS(fit_preds, file.path(plotdir, "COVSPECTRUM fitted lineage frequencies global multinomial fit.rds"))
# PLOT MULTINOMIAL FIT ON LOGIT SCALE ####
ncls = round(sqrt(length(unique(data$division))))
pl = qplot(data=fit_preds[fit_preds$variant!="Other",],
x=date, y=predicted, geom="blank") +
facet_wrap(~ division, ncol=ncls) +
geom_ribbon(aes(y=predicted, ymin=conf.low, ymax=conf.high, colour=NULL,
fill=variant), alpha=I(0.3)) +
geom_line(aes(colour=variant), alpha=I(1)) +
ylab("Share among newly diagnosed infections (%)") +
theme_hc() + xlab("") +
ggtitle("SARS-CoV2 LINEAGE FREQUENCIES",
subtitle=subtit) +
scale_x_date(date_breaks = brks, date_labels = lbls) +
theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
scale_y_continuous( trans="logit", breaks=c(10^seq(-5,0),0.5,0.9,0.99,0.999),
labels = c("0.001","0.01","0.1","1","10","100","50","90","99","99.9")) +
scale_fill_manual("variant", values=tail(as.vector(lineage_cols),-1)) +
scale_colour_manual("variant", values=tail(as.vector(lineage_cols),-1)) +
geom_point(data=data_agbyweekcountry1[data_agbyweekcountry1$variant!="Other",],
aes(x=date, y=prop, size=total,
colour=variant
),
alpha=I(1)) +
scale_size_continuous("total number\nsequenced", trans="sqrt",
range=c(0.1, 3), limits=c(1,max(data_agbyweekcountry1$total)),
breaks=c(10,100,1000, 10000)) +
# guides(fill=FALSE) +
# guides(colour=FALSE) +
theme(legend.position = "right") +
xlab("Collection date (start of week)") +
coord_cartesian(xlim=c(min(fit_preds$date), NA), # c(as.Date("2021-01-01"),NA),
ylim=c(0.0001, 0.99901), expand=0) +
theme(plot.title=element_text(size=25)) +
theme(plot.subtitle=element_text(size=20)) +
labs(tag = tag) + theme(plot.tag.position = "bottomright", plot.tag = element_text(vjust = 1, hjust = 1, size=8)) # +
# theme(strip.text.x = element_text(size = 20))
pl
if (ncls!=2) ggsave(file=file.path(plotdir, "predicted lineage freqs_logit scale.png"),
width=3*ncls,
height=(3/1.5)*ncls) else ggsave(file=file.path(plotdir, "predicted lineage freqs_logit scale.png"),
width=5*ncls,
height=4*ncls)
if (source!="COGUK") {
# zoomed in on last 6 months
pl = qplot(data=fit_preds[fit_preds$variant!="Other",],
x=date, y=predicted, geom="blank") +
facet_wrap(~ division) +
geom_ribbon(aes(y=predicted, ymin=conf.low, ymax=conf.high, colour=NULL,
fill=variant), alpha=I(0.3)) +
geom_line(aes(colour=variant), alpha=I(1)) +
ylab("Share among newly diagnosed infections (%)") +
theme_hc() + xlab("") +
ggtitle("SARS-CoV2 LINEAGE FREQUENCIES",
subtitle=paste0("GISAID data up to ",today, ", multinomial fit \n", model, ",\nall countries with >=", minseqs, " XBB.1.5* sequences included, using NextCladePangolin lineages")) +
scale_x_date(date_breaks = "1 month", date_labels = "%b %Y", expand=FALSE) +
theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
scale_y_continuous( trans="logit", breaks=c(10^seq(-5,0),0.5,0.9,0.99,0.999),
labels = c("0.001","0.01","0.1","1","10","100","50","90","99","99.9")) +
scale_fill_manual("variant", values=tail(as.vector(lineage_cols),-1)) +
scale_colour_manual("variant", values=tail(as.vector(lineage_cols),-1)) +
geom_point(data=data_agbyweekcountry1[data_agbyweekcountry1$variant!="Other",],
aes(x=date, y=prop, size=total,
colour=variant
),
alpha=I(1)) +
scale_size_continuous("total number\nsequenced", trans="sqrt",
range=c(0.1, 3), limits=c(1,max(data_agbyweekcountry1$total)),
breaks=c(10,100,1000, 10000)) +
# guides(fill=FALSE) +
# guides(colour=FALSE) +
theme(legend.position = "right") +
xlab("Collection date (start of week)") +
coord_cartesian(xlim=c(today-30*6,NA),
ylim=c(0.0001, 0.99901), expand=0) +
labs(tag = tag) +
theme(plot.tag.position = "bottomright",
plot.tag = element_text(vjust = 1, hjust = 1, size=8)) +
theme(plot.title=element_text(size=25)) +
theme(plot.subtitle=element_text(size=20))
pl
ggsave(file=file.path(plotdir, "predicted lineage freqs_last6months_logit scale.png"),
width=3*ncls,
height=(3/1.5)*ncls)
# plot with predictions per division/country for each region/continent
regions = unique(fit_preds$region)
for (region in regions) {
datsubs = fit_preds[fit_preds$variant!="Other"&fit_preds$region==region,]
ndivisions = length(unique(datsubs$division))
pl = qplot(data=datsubs,
x=date, y=predicted, geom="blank") +
facet_wrap(~ division, ncol=1) +
geom_ribbon(aes(y=predicted, ymin=conf.low, ymax=conf.high, colour=NULL,
fill=variant), alpha=I(0.3)) +
geom_line(aes(colour=variant), alpha=I(1)) +
ylab("Share among newly diagnosed infections (%)") +
theme_hc() + xlab("") +
ggtitle(paste0("SARS-CoV2 LINEAGE FREQUENCIES IN ", toupper(region)),
subtitle=subtit) +
theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
scale_y_continuous( trans="logit", breaks=c(10^seq(-5,0),0.5,0.9,0.99,0.999),
labels = c("0.001","0.01","0.1","1","10","100","50","90","99","99.9")) +
scale_fill_manual("", values=tail(as.vector(lineage_cols),-1)) +
scale_colour_manual("", values=tail(as.vector(lineage_cols),-1)) +
geom_point(data=data_agbyweekcountry1[data_agbyweekcountry1$variant!="Other"&data_agbyweekcountry1$region==region,],
aes(x=date, y=prop, size=total,
colour=variant
),
alpha=I(1)) +
scale_size_continuous("total number\nsequenced", trans="sqrt",
range=c(0.5, 4), limits=c(1,max(data_agbyweekcountry1$total)),
breaks=c(10,100,1000, 10000), guide=F) +
# guides(fill=FALSE) +
# guides(colour=FALSE) +
theme(legend.position = "bottom") +
xlab("Collection date (start of week)") +
coord_cartesian(xlim=c(as.Date("2021-01-01"),NA),
ylim=c(0.0001, 0.99901), expand=0) +
labs(tag = tag) +
theme(plot.tag.position = "bottomright",
plot.tag = element_text(vjust = 1, hjust = 1, size=8)) +
scale_x_date(date_breaks = brks, date_labels = lbls)
# theme(plot.title=element_text(size=40))
# theme(plot.subtitle=element_text(size=20))
pl
ggsave(file=file.path(plotdir, paste0("predicted lineage freqs_", region, "_logit scale.png")), width=9, height=3+2*ndivisions)
}
}
# separate plots for each division/country
divisions = unique(data_agbyweekcountry1$division)
for (division in divisions) {
# plot just for given division
pl = qplot(data=fit_preds[fit_preds$variant!="Other"&fit_preds$division==division,],
x=date, y=predicted, geom="blank") +
# facet_wrap(~ country) +
geom_ribbon(aes(y=predicted, ymin=conf.low, ymax=conf.high, colour=NULL,
fill=variant), alpha=I(0.3)) +
geom_line(aes(colour=variant), alpha=I(1)) +
ylab("Share among newly diagnosed\ninfections (%)") +
theme_hc() + xlab("") +
ggtitle(paste0("SARS-CoV2 LINEAGE FREQUENCIES IN ", toupper(division)),
subtitle=subtit) +
scale_x_date(date_breaks = brks, date_labels = lbls) +
theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
scale_y_continuous( trans="logit", breaks=c(10^seq(-5,0),0.5,0.9,0.99,0.999),
labels = c("0.001","0.01","0.1","1","10","100","50","90","99","99.9")) +
scale_fill_manual("", values=tail(as.vector(lineage_cols),-1)) +
scale_colour_manual("", values=tail(as.vector(lineage_cols),-1)) +
geom_point(data=data_agbyweekcountry1[data_agbyweekcountry1$variant!="Other"&
data_agbyweekcountry1$division==division,],
aes(x=date, y=prop, size=total,
colour=variant
),
alpha=I(1)) +
scale_size_continuous("total number\nsequenced", trans="sqrt",
range=c(0.5, 4), limits=c(1,max(data_agbyweekcountry1$total)),
breaks=c(10,100,1000, 10000), guide=F) +
# guides(fill=FALSE) +
# guides(colour=FALSE) +
theme(legend.position = "bottom") +
xlab("Collection date (start of week)") +
coord_cartesian(xlim=c(min(fit_preds$date), NA), # c(as.Date("2021-01-01"),NA),
ylim=c(0.0001, 0.99901), expand=0) +
labs(tag = tag) +
theme(plot.tag.position = "bottomright",
plot.tag = element_text(vjust = 1, hjust = 1, size=8)) # +
# theme(plot.title=element_text(size=25)) +
# theme(plot.subtitle=element_text(size=20))
pl
ggsave(file=file.path(plotdir, paste0("predicted lineage freqs_",division,"_logit scale.png")), width=9, height=6)
}
# # STILL NEED TO FINISH PART BELOW ####
#
# # map variant shares onto estimated infections as estimated by IHME ####
#
# ihme = bind_rows(read_csv("https://ihmecovid19storage.blob.core.windows.net/archive/2022-12-16/data_download_file_reference_2020.csv"),
# read_csv("https://ihmecovid19storage.blob.core.windows.net/archive/2022-12-16/data_download_file_reference_2021.csv"),
# read_csv("https://ihmecovid19storage.blob.core.windows.net/archive/2022-12-16/data_download_file_reference_2022.csv"),
# read_csv("https://ihmecovid19storage.blob.core.windows.net/archive/2022-12-16/data_download_file_reference_2023.csv"))
# names(ihme)
# sort(unique(ihme$location_name))
#
# # for England: map variant shares onto incidence data derived from ONS prevalence data
# # see https://github.com/epiforecasts/inc2prev/tree/master/data-processed
#
#
# # map US variant shares by region onto cases & wastewater surveillance data, cf. github Biobot Analytics ####
# # https://github.com/biobotanalytics/covid19-wastewater-data
# # I will need to estimate incidence from wastewater surveillance prevalence data then
# wastewater_US_byregion = read.csv("https://raw.githubusercontent.com/biobotanalytics/covid19-wastewater-data/master/wastewater_by_region.csv") %>%
# rename(date = sampling_week,
# division = region) %>%
# select(-population) %>%
# filter(division != "Nationwide")
# wastewater_US_byregion$date = as.Date(wastewater_US_byregion$date)
#
# # # linearly interpolate concentrations for all weekdays
# # grid = expand.grid(division=unique(wastewater_US_byregion$division),
# # date=seq(min(wastewater_US_byregion$date),
# # today, by = '1 day') )
# # pop_by_year_interpol = left_join(grid, pop_by_year) %>%
# # mutate(POPULATION =
# # interp1(x=as.numeric(DATE)[!is.na(POPULATION)],
# # y=POPULATION[!is.na(POPULATION)],
# # xi=as.numeric(DATE),
# # method="linear", # or cubic or spline
# # extrap=TRUE) )
#
#
# wastewater_US_byregion$division = paste0("USA - ", wastewater_US_byregion$division)
# wastewater_US_byregion$division = factor(wastewater_US_byregion$division, levels=as.character(levels(fit_preds$division)))
#
# wastewater_US_bycounty = read.csv("https://raw.githubusercontent.com/biobotanalytics/covid19-wastewater-data/master/wastewater_by_county.csv") %>%
# rename(date = sampling_week) %>%
# select(-X)
# wastewater_US_bycounty$date = as.Date(wastewater_US_bycounty$date)
#
# fit_preds$effective_concentration_rolling_average = NULL
# fit_preds = fit_preds %>% left_join(wastewater_US_byregion, by=c("date","division"))
#
#
# cases_US_byregion = read.csv("https://raw.githubusercontent.com/biobotanalytics/covid19-wastewater-data/master/cases_by_region.csv")
#
# cases_US_bycounty = read.csv("https://raw.githubusercontent.com/biobotanalytics/covid19-wastewater-data/master/cases_by_county.csv")
#
#
# # TO DO: DIDN'T UPDATE PART BELOW YET
# # (map share of variants on cases & hospitalisations etc - should get US data by state)
# # TO DO: map variant share onto US Biobot wastewater data
# # https://github.com/biobotanalytics/covid19-wastewater-data
#
#
# # plot predicted values as Muller plot
# pl = ggplot(data=fit_preds[fit_preds$date>=as.Date("2021-01-01"),],
# aes(x=date, y=predicted, group=variant)) +
# facet_wrap(~ division, ncol=ncls) +
# geom_area(aes(width=I(10), colour=NULL, fill=variant, group=variant), position="fill") +
# scale_fill_manual("", values=lineage_cols) +
# scale_x_date(date_breaks = "3 months", date_labels = "%b %Y") +
# theme(axis.text.x = element_text(angle = 60, vjust = 1, hjust=1)) +
# # guides(color = guide_legend(reverse=F, nrow=2, byrow=T), fill = guide_legend(reverse=F, nrow=2, byrow=T)) +
# theme_hc() +
# ylab("Share") +
# theme(legend.position="right",
# axis.title.x=element_blank()) +
# labs(title = "SARS-CoV2 LINEAGE FREQUENCIES",
# subtitle=paste0("GISAID data up to ",today, ", multinomial fit ", model, ",\nall countries with >=", minseqs, " XBB.1.5* sequences shown")) +
# annotate("rect",
# xmin=max(data$date)+1,
# xmax=as.Date(date.to, origin="1970-01-01")+5,
# ymin=0, ymax=1, alpha=0.5, fill="white") + # extrapolated part
# theme(plot.title=element_text(size=25)) +
# theme(plot.subtitle=element_text(size=20))
# pl
#
# ggsave(file=file.path(plotdir, "muller plot.png"),
# width=4*ncls,
# height=(4/1.2)*ncls)
#
#
# # MAP VARIANT SHARE ONTO CASE NUMBERS ####
#