Estimating the prevalence of two or more diseases using outcomes from multiplex group testing.

When screening a population for infectious diseases, pooling individual specimens (e.g., blood, swabs, urine, etc.) can provide enormous cost savings when compared to testing specimens individually. In the biostatistics literature, testing pools of specimens is commonly known as group testing or pooled testing. Although estimating a population-level prevalence with group testing data has received a large amount of attention, most of this work has focused on applications involving a single disease, such as human immunodeficiency virus. Modern methods of screening now involve testing pools and individuals for multiple diseases simultaneously through the use of multiplex assays. Hou et al. (2017, Biometrics, 73, 656-665) and Hou et al. (2020, Biostatistics, 21, 417-431) recently proposed group testing protocols for multiplex assays and derived relevant case identification characteristics, including the expected number of tests and those which quantify classification accuracy. In this article, we describe Bayesian methods to estimate population-level disease probabilities from implementing these protocols or any other multiplex group testing protocol which might be carried out in practice. Our estimation methods can be used with multiplex assays for two or more diseases while incorporating the possibility of test misclassification for each disease. We use chlamydia and gonorrhea testing data collected at the State Hygienic Laboratory at the University of Iowa to illustrate our work. We also provide an online R resource practitioners can use to implement the methods in this article.

binGroup2: Statistical Tools for Infection Identification via Group Testing.

Group testing is the process of testing items as an amalgamation, rather than separately, to determine the binary status for each item. Its use was especially important during the COVID-19 pandemic through testing specimens for SARS-CoV-2. The adoption of group testing for this and many other applications is because members of a negative testing group can be declared negative with potentially only one test. This subsequently leads to significant increases in laboratory testing capacity. Whenever a group testing algorithm is put into practice, it is critical for laboratories to understand the algorithm's operating characteristics, such as the expected number of tests. Our paper presents the binGroup2 package that provides the statistical tools for this purpose. This R package is the first to address the identification aspect of group testing for a wide variety of algorithms. We illustrate its use through COVID-19 and chlamydia/gonorrhea applications of group testing.

Informative array testing with multiplex assays.

High-volume testing of clinical specimens for sexually transmitted diseases is performed frequently by a process known as group testing. This algorithmic process involves testing portions of specimens from separate individuals together as one unit (or "group") to detect diseases. Retesting is performed on groups that test positively in order to differentiate between positive and negative individual specimens. The overall goal is to use the least number of tests possible across all individuals without sacrificing diagnostic accuracy. One of the most efficient group testing algorithms is array testing. In its simplest form, specimens are arranged into a grid-like structure so that row and column groups can be formed. Positive-testing rows/columns indicate which specimens to retest. With the growing use of multiplex assays, the increasing number of diseases tested by these assays, and the availability of subject-specific risk information, opportunities exist to make this testing process even more efficient. We propose specific specimen arrangements within an array that can reduce the number of retests needed when compared with other array testing algorithms. We examine how to calculate operating characteristics, including the expected number of tests and the SD for the number of tests, and then subsequently find a best arrangement. Our methods are illustrated for chlamydia and gonorrhea detection with the Aptima Combo 2 Assay. We also provide R functions to make our research accessible to laboratories.

Incorporating the dilution effect in group testing regression.

When screening for infectious diseases, group testing has proven to be a cost efficient alternative to individual level testing. Cost savings are realized by testing pools of individual specimens (eg, blood, urine, saliva, and so on) rather than by testing the specimens separately. However, a common concern that arises in group testing is the so-called "dilution effect." This occurs if the signal from a positive individual's specimen is diluted past an assay's threshold of detection when it is pooled with multiple negative specimens. In this article, we propose a new statistical framework for group testing data that merges estimation and case identification, which are often treated separately in the literature. Our approach considers analyzing continuous biomarker levels (eg, antibody levels, antigen concentrations, and so on) from pooled samples to estimate both a binary regression model for the probability of disease and the biomarker distributions for cases and controls. To increase case identification accuracy, we then show how estimates of the biomarker distributions can be used to select diagnostic thresholds on a pool-by-pool basis. Our proposals are evaluated through numerical studies and are illustrated using hepatitis B virus data collected on a prison population in Ireland.

Generalized additive regression for group testing data.

In screening applications involving low-prevalence diseases, pooling specimens (e.g., urine, blood, swabs, etc.) through group testing can be far more cost effective than testing specimens individually. Estimation is a common goal in such applications and typically involves modeling the probability of disease as a function of available covariates. In recent years, several authors have developed regression methods to accommodate the complex structure of group testing data but often under the assumption that covariate effects are linear. Although linearity is a reasonable assumption in some applications, it can lead to model misspecification and biased inference in others. To offer a more flexible framework, we propose a Bayesian generalized additive regression approach to model the individual-level probability of disease with potentially misclassified group testing data. Our approach can be used to analyze data arising from any group testing protocol with the goal of estimating multiple unknown smooth functions of covariates, standard linear effects for other covariates, and assay classification accuracy probabilities. We illustrate the methods in this article using group testing data on chlamydia infection in Iowa.

More powerful goodness-of-fit tests for uniform stochastic ordering.

The ordinal dominance curve (ODC) is a useful graphical tool to compare two population distributions. These distributions are said to satisfy uniform stochastic ordering (USO) if the ODC for them is star-shaped. A goodness-of-fit test for USO was recently proposed when both distributions are unknown. This test involves calculating the L p distance between an empirical estimator of the ODC and its least star-shaped majorant. The least favorable configuration of the two distributions was established so that proper critical values could be determined; i.e., to control the probability of type I error for all star-shaped ODCs. However, the use of these critical values can lead to a conservative test and minimal power to detect certain non-star-shaped alternatives. Two new methods for determining data-dependent critical values are proposed. Simulation is used to show both methods can provide substantial increases in power while still controlling the size of the distance-based test. The methods are also applied to a data set involving premature infants. An R package that implements all tests is provided.

Tests in short supply? Try group testing.

Christopher R. Bilder, Peter C. Iwen, Baha Abdalhamid, Joshua M. Tebbs and Christopher S. McMahan explain how, by pooling specimens, testing capacity for SARS-CoV-2 can be increased.

Array testing for multiplex assays.

Group testing involves pooling individual specimens (e.g., blood, urine, swabs, etc.) and testing the pools for the presence of disease. When the proportion of diseased individuals is small, group testing can greatly reduce the number of tests needed to screen a population. Statistical research in group testing has traditionally focused on applications for a single disease. However, blood service organizations and large-scale disease surveillance programs are increasingly moving towards the use of multiplex assays, which measure multiple disease biomarkers at once. Tebbs and others (2013, Two-stage hierarchical group testing for multiple infections with application to the Infertility Prevention Project. Biometrics69, 1064-1073) and Hou and others (2017, Hierarchical group testing for multiple infections. Biometrics73, 656-665) were the first to examine hierarchical group testing case identification procedures for multiple diseases. In this article, we propose new non-hierarchical procedures which utilize two-dimensional arrays. We derive closed-form expressions for the expected number of tests per individual and classification accuracy probabilities and show that array testing can be more efficient than hierarchical procedures when screening individuals for multiple diseases at once. We illustrate the potential of using array testing in the detection of chlamydia and gonorrhea for a statewide screening program in Iowa. Finally, we describe an R/Shiny application that will help practitioners identify the best multiple-disease case identification algorithm.

From mixed effects modeling to spike and slab variable selection: A Bayesian regression model for group testing data.

Due to reductions in both time and cost, group testing is a popular alternative to individual-level testing for disease screening. These reductions are obtained by testing pooled biospecimens (eg, blood, urine, swabs, etc.) for the presence of an infectious agent. However, these reductions come at the expense of data complexity, making the task of conducting disease surveillance more tenuous when compared to using individual-level data. This is because an individual's disease status may be obscured by a group testing protocol and the effect of imperfect testing. Furthermore, unlike individual-level testing, a given participant could be involved in multiple testing outcomes and/or may never be tested individually. To circumvent these complexities and to incorporate all available information, we propose a Bayesian generalized linear mixed model that accommodates data arising from any group testing protocol, estimates unknown assay accuracy probabilities and accounts for potential heterogeneity in the covariate effects across population subgroups (eg, clinic sites, etc.); this latter feature is of key interest to practitioners tasked with conducting disease surveillance. To achieve model selection, our proposal uses spike and slab priors for both fixed and random effects. The methodology is illustrated through numerical studies and is applied to chlamydia surveillance data collected in Iowa.

The objective function controversy for group testing: Much ado about nothing?

Group testing is an indispensable tool for laboratories when testing high volumes of clinical specimens for infectious diseases. An important decision that needs to be made prior to implementation is determining what group sizes to use. In best practice, an objective function is chosen and then minimized to determine an optimal set of these group sizes, known as the optimal testing configuration (OTC). There are a few options for objective functions, and they differ based on how the expected number of tests, assay characteristics, and testing constraints are taken into account. These varied options have led to a recent controversy in the literature regarding which of two different objective functions is better. In our paper, we examine these objective functions over a number of realistic situations for infectious disease testing. We show that this controversy may be much ado about nothing because the OTCs and corresponding results (eg, number of tests and accuracy) are largely the same for standard testing algorithms in a wide variety of situations.

Testing for positive quadrant dependence.

We develop an empirical likelihood approach to test independence of two univariate random variables X and Y versus the alternative that X and Y are strictly positive quadrant dependent (PQD). Establishing this type of ordering between X and Y is of interest in many applications, including finance, insurance, engineering, and other areas. Adopting the framework in Einmahl and McKeague (2003, Bernoulli), we create a distribution-free test statistic that integrates a localized empirical likelihood ratio test statistic with respect to the empirical joint distribution of X and Y. When compared to well known existing tests and distance-based tests we develop by using copula functions, simulation results show the EL testing procedure performs well in a variety of scenarios when X and Y are strictly PQD. We use three data sets for illustration and provide an online R resource practitioners can use to implement the methods in this article.

Informative group testing for multiplex assays.

Infectious disease testing frequently takes advantage of two tools-group testing and multiplex assays-to make testing timely and cost effective. Until the work of Tebbs et al. (2013) and Hou et al. (2017), there was no research available to understand how best to apply these tools simultaneously. This recent work focused on applications where each individual is considered to be identical in terms of the probability of disease. However, risk-factor information, such as past behavior and presence of symptoms, is very often available on each individual to allow one to estimate individual-specific probabilities. The purpose of our paper is to propose the first group testing algorithms for multiplex assays that take advantage of individual risk-factor information as expressed by these probabilities. We show that our methods significantly reduce the number of tests required while preserving accuracy. Throughout this paper, we focus on applying our methods with the Aptima Combo 2 Assay that is used worldwide for chlamydia and gonorrhea screening.

Group testing case identification with biomarker information.

Screening procedures for infectious diseases, such as HIV, often involve pooling individual specimens together and testing the pools. For diseases with low prevalence, group testing (or pooled testing) can be used to classify individuals as diseased or not while providing considerable cost savings when compared to testing specimens individually. The pooling literature is replete with group testing case identification algorithms including Dorfman testing, higher-stage hierarchical procedures, and array testing. Although these algorithms are usually evaluated on the basis of the expected number of tests and classification accuracy, most evaluations in the literature do not account for the continuous nature of the testing responses and thus invoke potentially restrictive assumptions to characterize an algorithm's performance. Commonly used case identification algorithms in group testing are considered and are evaluated by taking a different approach. Instead of treating testing responses as binary random variables (i.e., diseased/not), evaluations are made by exploiting an assay's underlying continuous biomarker distributions for positive and negative individuals. In doing so, a general framework to describe the operating characteristics of group testing case identification algorithms is provided when these distributions are known. The methodology is illustrated using two HIV testing examples taken from the pooling literature.

Group testing regression models with dilution submodels.

Group testing, where specimens are tested initially in pools, is widely used to screen individuals for sexually transmitted diseases. However, a common problem encountered in practice is that group testing can increase the number of false negative test results. This occurs primarily when positive individual specimens within a pool are diluted by negative ones, resulting in positive pools testing negatively. If the goal is to estimate a population-level regression model relating individual disease status to observed covariates, severe bias can result if an adjustment for dilution is not made. Recognizing this as a critical issue, recent binary regression approaches in group testing have utilized continuous biomarker information to acknowledge the effect of dilution. In this paper, we have the same overall goal but take a different approach. We augment existing group testing regression models (that assume no dilution) with a parametric dilution submodel for pool-level sensitivity and estimate all parameters using maximum likelihood. An advantage of our approach is that it does not rely on external biomarker test data, which may not be available in surveillance studies. Furthermore, unlike previous approaches, our framework allows one to formally test whether dilution is present based on the observed group testing data. We use simulation to illustrate the performance of our estimation and inference methods, and we apply these methods to 2 infectious disease data sets.

Bayesian regression for group testing data.

Group testing involves pooling individual specimens (e.g., blood, urine, swabs, etc.) and testing the pools for the presence of a disease. When individual covariate information is available (e.g., age, gender, number of sexual partners, etc.), a common goal is to relate an individual's true disease status to the covariates in a regression model. Estimating this relationship is a nonstandard problem in group testing because true individual statuses are not observed and all testing responses (on pools and on individuals) are subject to misclassification arising from assay error. Previous regression methods for group testing data can be inefficient because they are restricted to using only initial pool responses and/or they make potentially unrealistic assumptions regarding the assay accuracy probabilities. To overcome these limitations, we propose a general Bayesian regression framework for modeling group testing data. The novelty of our approach is that it can be easily implemented with data from any group testing protocol. Furthermore, our approach will simultaneously estimate assay accuracy probabilities (along with the covariate effects) and can even be applied in screening situations where multiple assays are used. We apply our methods to group testing data collected in Iowa as part of statewide screening efforts for chlamydia, and we make user-friendly R code available to practitioners.

NONPARAMETRIC GOODNESS-OF-FIT TESTS FOR UNIFORM STOCHASTIC ORDERING.

We propose Lp distance-based goodness-of-fit (GOF) tests for uniform stochastic ordering with two continuous distributions F and G, both of which are unknown. Our tests are motivated by the fact that when F and G are uniformly stochastically ordered, the ordinal dominance curve R = FG-1 is star-shaped. We derive asymptotic distributions and prove that our testing procedure has a unique least favorable configuration of F and G for p ∈ [1,∞]. We use simulation to assess finite-sample performance and demonstrate that a modified, one-sample version of our procedure (e.g., with G known) is more powerful than the one-sample GOF test suggested by Arcones and Samaniego (2000, Annals of Statistics). We also discuss sample size determination. We illustrate our methods using data from a pharmacology study evaluating the effects of administering caffeine to prematurely born infants.

Hierarchical group testing for multiple infections.

Group testing, where individuals are tested initially in pools, is widely used to screen a large number of individuals for rare diseases. Triggered by the recent development of assays that detect multiple infections at once, screening programs now involve testing individuals in pools for multiple infections simultaneously. Tebbs, McMahan, and Bilder (2013, Biometrics) recently evaluated the performance of a two-stage hierarchical algorithm used to screen for chlamydia and gonorrhea as part of the Infertility Prevention Project in the United States. In this article, we generalize this work to accommodate a larger number of stages. To derive the operating characteristics of higher-stage hierarchical algorithms with more than one infection, we view the pool decoding process as a time-inhomogeneous, finite-state Markov chain. Taking this conceptualization enables us to derive closed-form expressions for the expected number of tests and classification accuracy rates in terms of transition probability matrices. When applied to chlamydia and gonorrhea testing data from four states (Region X of the United States Department of Health and Human Services), higher-stage hierarchical algorithms provide, on average, an estimated 11% reduction in the number of tests when compared to two-stage algorithms. For applications with rarer infections, we show theoretically that this percentage reduction can be much larger.

Estimating the prevalence of multiple diseases from two-stage hierarchical pooling.

Testing protocols in large-scale sexually transmitted disease screening applications often involve pooling biospecimens (e.g., blood, urine, and swabs) to lower costs and to increase the number of individuals who can be tested. With the recent development of assays that detect multiple diseases, it is now common to test biospecimen pools for multiple infections simultaneously. Recent work has developed an expectation-maximization algorithm to estimate the prevalence of two infections using a two-stage, Dorfman-type testing algorithm motivated by current screening practices for chlamydia and gonorrhea in the USA. In this article, we have the same goal but instead take a more flexible Bayesian approach. Doing so allows us to incorporate information about assay uncertainty during the testing process, which involves testing both pools and individuals, and also to update information as individuals are tested. Overall, our approach provides reliable inference for disease probabilities and accurately estimates assay sensitivity and specificity even when little or no information is provided in the prior distributions. We illustrate the performance of our estimation methods using simulation and by applying them to chlamydia and gonorrhea data collected in Nebraska. Copyright © 2016 John Wiley & Sons, Ltd.