Applications of PCR (Real-Time and MassTag) and Enzyme-Linked Immunosorbent Assay in Diagnosis of Respiratory Infections and Diarrheal Illness Among Deployed U.S. Military Personnel During Exercise Balikatan 2009, Philippines

Abstract

Laboratory-based surveillance for diarrheal and respiratory illness was conducted at the 2009 Republic of the Philippines-United States Balikatan exercise to determine the presence of specific pathogens endemic in the locations where the military exercises were conducted. Ten stool and 6 respiratory specimens were obtained from individuals meeting case definitions for diarrhea or respiratory illness. Stool specimens were frozen in dry ice and remotely tested using enzyme-linked immunosorbent assay for Rotavirus, Astrovirus, Adenovirus, Entamoeba histolytica, Giardia, and Cryptosporidium and polymerase chain reaction for enterotoxigenic Escherichia coli, Campylobacter, Shigella, Vibrio, Salmonella, and Norovirus. Eight (4 for Campylobacter jejuni, 2 for Campylobacter coli, 1 for Norovirus genogroup II, and 1 for both Campylobacter coli and enterotoxigenic Escherichia coli) of 10 samples were positive for at least 1 enteric pathogen. MassTag polymerase chain reaction for influenza A and B, respiratory syncytial virus groups A and B, human coronavirus-229E and human coronavirus-OC43, human metapneumovirus, enterovirus, human parainfluenza viruses 2,3, and 4a, human adenovirus, Haemophilus influenzae, Neisseria meningitidis, Streptococcus pneumoniae, Legionella pneumonia, and Mycoplasma pneumonia was done on respiratory specimens. Out of 6 samples, 3 tested positive for H. influenzae; 1 tested positive for both H. influenzae and human parainfluenza virus 3; and 2 tested negative. Laboratory-based surveillance can be useful in determining etiologies of diarrheal and respiratory illness of deployed military personnel.

Practical Guidance for Clinical Microbiology Laboratories: Laboratory Diagnosis of Parasites from the Gastrointestinal Tract

“…Methods that have been used for differentiation of E. histolytica from other amoebae include PCR-restriction fragment length polymorphism (RFLP) analysis and real-time PCR with either SYBR green or specific probes (230-232). The use of highly sensitive tests that differentiate E. histolytica from E. dispar and other morphologically similar amoebae has allowed us to refine the determination of the prevalence and incidence of infections, to detect the presence of the organism in zoonotic hosts, to discover uncommon modes of transmission, and to study the epidemiology of the disease (233)(234)(235)(236)(237). Not surprisingly, the prevalence of infection by E. histolytica has been demonstrated to be lower than previously thought, since the methods used (i.e., microscopy and enzyme immunoassay) mischaracterized E. dispar as E. histolytica (238).…”

Reverse Transcription Polymerase Chain Reaction (RT-PCR) as a Tool for Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) Surveillance in the Military

“…Although RT-PCR has a high sensitivity and reliability, the procedure may disrupt the viral genetic material, causing false-negative results in some cases (43). The outbreaks of acute respiratory illnesses are often considered health priorities during military postings, deployments, and operations (44). Influenza and acute respiratory infections can significantly impair military operations (44) and can be used as a pathogen(s) or bio-warfare to suppress enemies during war confrontations.…”

“…The outbreaks of acute respiratory illnesses are often considered health priorities during military postings, deployments, and operations (44). Influenza and acute respiratory infections can significantly impair military operations (44) and can be used as a pathogen(s) or bio-warfare to suppress enemies during war confrontations. The value of RT-PCR lies in its ability to detect RNA viruses in a given specimen, amplify target genes, and measure viral loads or monitor responses to antiviral drugs (45).…”

“…Observing good personal hygiene (washing hands and covering nose when sneezing and coughing), sanitizing public utilities such as door handles, stair rails, as well as providing health education to armed forces may help to contain such diseases in military organizations (47). Laboratory-based surveillance often plays a key role in disease prevention via detecting sources of pathogens, contributing to a more reliable risk assessment of the infectious disease (44,48). Such knowledge can also be useful in formulating successful prevention methods and providing care to deployed military personnel.…”