Diagnostic Challenges of Typhoid Fever

Unmasking the Diagnostic Challenges of Typhoid Fever

Diagnostic Challenges of Typhoid Fever: Typhoid fever, caused by the bacterium Salmonella typhi, has been a persistent global health challenge for centuries. Despite advances in medical science, this infectious disease continues to pose diagnostic challenges that hinder timely treatment and the prevention of its spread. In this article, we delve into the intricacies of diagnosing typhoid fever, exploring the hurdles and potential solutions to improve diagnostic accuracy.

The Complexity of Typhoid Fever Diagnosis

Diagnostic Challenges of Typhoid Fever is far from straightforward, primarily due to several factors:

  1. Nonspecific Symptoms: Typhoid fever often presents with vague, nonspecific symptoms that overlap with many other common illnesses, including fever, headache, malaise, abdominal pain, and diarrhea. This makes clinical diagnosis challenging, as these symptoms are not unique to typhoid.
  2. Limited Sensitivity of Traditional Tests: Traditional diagnostic methods like blood culture, which involve isolating the bacterium from a patient’s blood, are considered the gold standard. However, they are time-consuming, costly, and lack the sensitivity to detect the bacteria in the early stages of infection.
  3. Asymptomatic Carriers: Typhoid carriers are individuals who harbour the Salmonella typhi bacteria without displaying any symptoms of the disease themselves. After recovering from a typhoid fever episode, a small percentage of individuals continue to carry the bacteria in their intestinal tracts or gallbladders and shed them in their feces. These individuals are termed “chronic carriers.”

There are two types of carriers:

  • Temporary or Convalescent Carriers: These individuals excrete the bacteria for a short duration, usually a few weeks to a few months, after recovering from the acute phase of the disease.
  • Chronic Carriers: These individuals continue to excrete the bacteria for more than a year after recovery. In some cases, they may carry the bacteria for life.

The most famous example of a chronic carrier is “Typhoid Mary” (Mary Mallon), a cook in the early 20th century in the U.S. who was asymptomatic but spread the disease to many individuals through the food she prepared.


Diagnostic Challenge with Typhoid Carriers:

  • Asymptomatic Nature: Since carriers don’t show symptoms of the disease, they are often unaware that they are carriers. This makes it challenging to identify them unless there’s a specific reason to suspect and test for carrier status, such as tracing the source of an outbreak.
  • Intermittent Shedding: Carriers may not consistently shed the bacteria in their feces. So, a single negative stool culture does not necessarily rule out carrier status.
  • Low Bacterial Load: The amount of bacteria shed by carriers might be lower than in acutely infected patients. This makes detection in stool samples more difficult.
  • Need for Multiple Tests: To confirm carrier status, multiple stool samples might be required over time. This can be logistically challenging and may not be feasible in all settings.
  • Diagnostic Sensitivity: Standard methods like stool culture have limited sensitivity, especially in carriers. More advanced techniques like polymerase chain reaction (PCR) can be more sensitive but might not be available in all settings.
  1. Antibiotic Use: Prior use of antibiotics can suppress the growth of Salmonella typhi in blood cultures, leading to false-negative results. This is a common scenario as patients often seek medical attention and may be prescribed antibiotics before a definitive diagnosis is made.
  2. Geographical Variation: Typhoid fever incidence varies widely by region. Diagnostic challenges are compounded in areas with limited access to healthcare facilities and diagnostic tools.
  • Strain Variations: Different strains of Salmonella typhi might be prevalent in different regions. Some regions might also see a higher prevalence of related bacteria like Salmonella paratyphi, which causes paratyphoid fever.
  • Antibiotic Resistance: Certain regions, especially parts of Asia, have seen the emergence of multi-drug resistant (MDR) strains of Salmonella typhi. This has prompted increased vigilance and the need for antibiotic susceptibility testing.
  • Vaccination Coverage: In areas where typhoid vaccination is more prevalent, the incidence of the disease might be lower, influencing testing and detection strategies.
  • Endemic Areas: In areas where typhoid is endemic, clinical diagnosis based on symptoms might be more common, and empirical treatment might be initiated even before laboratory confirmation. This is because clinicians in these areas are more familiar with the presentation of the disease.
  • Laboratory Facilities: The availability and sophistication of laboratory facilities can vary. In some low-resource settings, advanced diagnostic tests might not be readily available, leading to reliance on clinical diagnosis or less sensitive tests.

6. Lack of Rapid Tests: While there are rapid diagnostic tests (RDTs) available for typhoid, their sensitivity and specificity can vary. Some may produce false-positive or false-negative results, adding to the diagnostic conundrum.

Here are some examples of typhoid RDTs and their associated issues:

Widal Test: This is one of the oldest and most commonly used tests in many parts of the world.


  • Low Specificity: It can cross-react with other febrile illnesses and other types of Salmonella.
  • Baseline Titers: Different regions may have different baseline antibody titers in the population, affecting interpretation.
  • Timing: The test is more reliable in the second week of illness, making early diagnosis challenging.

Paired Samples Needed: Ideally, both acute and convalescent-phase sera should be tested to demonstrate a significant rise in antibody titers.

Rapid Card Tests : They detect  IgM and IgG antibodies against a specific antigen of the Salmonella typhi bacterium.


  • Cross-Reactivity: There can be cross-reactivity with other Salmonella species.
  • Duration of Illness: IgM is usually detectable in the first week of fever, while IgG is detectable later. This differentiation is crucial, and the test might not always be reliable in differentiating between current and past infections.

Immunology Assays : These tests  use particle inhibition immunoassay to detect IgM antibodies against Salmonella typhi O9 lipopolysaccharide.


  • Cost: It’s more expensive than the Widal test, making it less accessible in low-resource settings.
  • Specificity and Sensitivity: While it’s generally considered more specific than the Widal test, its sensitivity and specificity can still vary based on the study and population.

Why do normal tests fail?

Traditional typhoid tests, specifically blood culture-based tests, can face limitations and may fail to provide accurate and timely diagnosis for several reasons:

Low Bacterial Load in Blood: During the early stages of typhoid fever, the concentration of Salmonella typhi bacteria in the bloodstream may be very low. Blood cultures rely on the ability to detect and isolate these bacteria, and when their numbers are minimal, it can result in false-negative results.

Interference from Prior Antibiotic Use: Patients who suspect they have typhoid fever often seek medical attention and may be prescribed antibiotics before a definitive diagnosis is made. Antibiotics can inhibit the growth of bacteria in blood cultures, making it difficult to detect Salmonella typhi/paratyphi.

Inadequate Sample Volume: Collecting an insufficient volume of blood for culture can also contribute to diagnostic failure. A small blood sample may not contain enough bacteria for detection.

Time-Consuming Process: Traditional blood culture tests typically take several days to produce results. This delay can be a significant drawback, as timely treatment is essential in managing typhoid fever effectively.

Laboratory Expertise: Blood culture tests require skilled laboratory personnel and well-equipped facilities. In resource-limited settings, where typhoid is prevalent, access to these resources may be limited, affecting the reliability of test results.

False Positives/Negatives: Blood culture is a critical diagnostic method for many bloodstream infections, including typhoid fever. However, the process from collection to result interpretation can be prone to errors, leading to false-positive or false-negative results. 

False Positives:

  • Contamination: This is the most common reason for false-positive blood cultures. Contamination can occur when skin microbes or environmental organisms are inadvertently introduced into the blood culture specimen during collection. Such contaminants can grow in the culture medium, suggesting the presence of a bloodstream infection when there isn’t one.
  • Examples: Common skin contaminants include coagulase-negative staphylococci, Propionibacterium acnes, and certain species of Corynebacterium. If these organisms are identified in a blood culture, it raises suspicion of contamination, especially if the patient’s clinical signs don’t align with a bloodstream infection.
  • Consequence: False-positive results can lead to unnecessary treatment with antibiotics, longer hospital stays, additional tests, increased costs, and potential antibiotic resistance development.

False Negatives:

  • Prior Antibiotic Administration: If a patient has received antibiotics before the blood sample collection, the number of bacteria in the bloodstream may be reduced, leading to a decreased likelihood of detecting the bacteria in culture.
  • Volume of Blood: The volume of the blood sample is critical. Insufficient blood can reduce the chances of detecting the bacteria. Conversely, too much blood can dilute the culture medium, making it less effective.
  • Frequency of Sampling: In some diseases, bacteremia (presence of bacteria in the blood) can be intermittent. Taking only a single blood culture can miss the bacteria, leading to false-negative results.
  • Culture Medium and Conditions: Not all bacteria grow well under standard blood culture conditions. If specific pathogens are suspected, specialized media or growth conditions might be needed.
  • Laboratory Errors: Mistakes in incubation time, temperature, or other conditions can hinder bacterial growth.
  • Consequence: False-negative results can delay appropriate treatment, prolonging illness and increasing the risk of complications. The patient might be wrongly reassured, and the opportunity for early intervention is missed.

Overall Impact: 

The consequences of both false-positive and false-negative blood cultures in clinical practice are significant. They can misguide clinical decisions, leading to either over-treatment or under-treatment, both of which can adversely affect patient outcomes. Therefore, ensuring proper blood culture techniques, from sample collection to result interpretation, is paramount for accurate diagnosis and appropriate patient management.

Advanced Stage and Complications:

 In untreated cases or cases that don’t respond to treatment, the disease might progress to more severe symptoms like intestinal perforation, bleeding, or neurological symptoms like delirium (termed “typhoid state”).

Even these severe symptoms can be mistaken for other conditions, like acute abdomen from different causes or central nervous system infections.

Other Overlapping Symptoms: Some patients with typhoid fever can present with a rash (rose spots) on the abdomen, cough, or joint pains. These manifestations are also seen in other diseases like rickettsial infections, viral exanthems, or other bacterial sepsis.

Variations Across Populations: The classic presentation of typhoid fever, as taught in medical textbooks, might not be the same across all populations or age groups. For instance, constipation is more commonly seen in adults, while children might present with diarrhea.

The course of the disease might be atypical in immunocompromised patients or those with other underlying conditions.

Endemic Areas vs. Non-endemic Areas: In areas where typhoid fever is endemic, clinicians might be more attuned to the nuances of the disease presentation and might suspect typhoid in febrile patients sooner. However, the flip side is that other febrile illnesses might be overlooked or misdiagnosed as typhoid. In non-endemic areas, typhoid might not be high on the list of differential diagnoses unless there’s a relevant travel history or exposure.

Potential Solutions and Advancements in the field of Diagnostic Challenges of Typhoid Fever

Efforts to improve typhoid fever diagnosis are ongoing. Promising developments include:

  1. Molecular Diagnostics: Advances in molecular biology have led to the development of Real Time Polymerase Chain Reaction (RT PCR) tests for typhoid. RT PCR offers increased sensitivity and specificity and can detect the bacterium’s DNA in blood or stool samples, even during early infection stages.
  2. Serological Tests: Enzyme-linked immunosorbent assay (ELISAs) that detect specific antibodies produced by the body in response to Salmonella typhi infection are being explored. These tests can help diagnose cases where blood cultures may yield false negatives.
  3. Improved RDTs: Researchers are working to enhance the accuracy of rapid diagnostic tests. These tests are especially valuable in resource-limited settings where timely diagnosis is crucial.
  4. Fecal Culture: Since Salmonella Typhi is also shed in feces, testing stool samples in addition to blood cultures can improve diagnostic accuracy, particularly in carriers.
  5. Public Health Measures: Combining diagnostic efforts with public health measures such as improved sanitation, access to clean water, and vaccination campaigns can reduce the burden of typhoid fever and its transmission.

Conclusion: Introduction to Vanscan

This diagnostic tool utilizes nucleic acid-based technology for pinpointing Typhoid, leveraging a proprietary method (Patent No. 355208). It analyzes DNA extracted from the blood of affected individuals, showcasing remarkable sensitivity and specificity.

The VANSCAN Typhoid RT PCR Test offers prompt and definitive identification of Typhoid fever, streamlining the selection of suitable antibiotics for treatment. This plays a pivotal role in combating the growing concern of Antimicrobial Resistance (AMR) in India.

The development of the VANSCAN Typhoid RT PCR Test stems from a collaborative effort between Vanguard Diagnostics and Graphic Era University, Dehradun.

Diagnostic Challenges of Typhoid Fever

How is VANSCAN revolutionizing Diagnostic challenges of Typhoid Fever, and how does it work?

  1. Sample Collection: To perform the assay, a biological sample from the patient is collected. Common sample types include blood, stool, or urine, depending on the stage of the infection and the type of assay used.
  2. Nucleic Acid Extraction: In this step, nucleic acids (DNA or RNA) are extracted from the patient’s sample. This process typically involves breaking open the bacterial cells (Salmonella Typhi in this case) and isolating their genetic material.
  3. Target Amplification: To increase the sensitivity of the assay, a technique called polymerase chain reaction (PCR) is often used. PCR selectively amplifies specific regions of the Salmonella Typhi genome if present in the patient’s sample. This step can make it easier to detect the bacterium’s genetic material, even if it’s present in low concentrations.
  4. Probe Hybridization: Nucleic acid probes, which are short pieces of DNA  designed to complement specific sequences of the Salmonella typhi genome, are used in the assay. These probes are labeled with a fluorescent or chemiluminescent marker. If Salmonella typhi DNA is present in the patient’s sample, the probes will bind (hybridize) to the matching sequences, forming a probe-target complex.
  5. Detection: The presence of the probe-target complex is detected using a specialized instrument that can measure the fluorescence emitted by the labeled probes. The intensity of the signal is directly proportional to the amount of Salmonella typhi genetic material in the sample.
  6. Interpretation: The assay results are interpreted based on the presence or absence of a detectable signal. Positive results indicate the presence of Salmonella typhi DNA  in the patient’s sample, suggesting an active infection.

Nucleic acid-based diagnostic assays offer several advantages for typhoid fever diagnosis:

High Sensitivity: These assays can detect low concentrations of Salmonella typhi genetic material, making them suitable for early infection detection.

Specificity: They are highly specific, as they target unique genetic sequences of the bacterium.

Rapid Results: This nucleic acid-based assays provide results within an hour, allowing for timely treatment decisions.

Less Affected by Antibiotic Use: Unlike culture-based methods, these assays are less affected by prior antibiotic use, as they don’t rely on bacterial growth.

Salmonella typhi and paratyphi. Unlike existing tests, such as Widal and rapid cards, VANSCAN offers superior sensitivity and specificity. By enabling early detection on day one of infection, this test facilitates timely administration of appropriate antibiotics, contributing significantly to combating Antimicrobial Resistance (AMR) in India. Developed through an industry-academia collaboration between Vanguard Diagnostics and Graphic Era University, Dehradun.