In this study, 810 deceased individuals referred to forensic medicine over six months in 2021, regardless of the cause of death, were included.
In our study, the highest number of deaths occurred at home, with 336 cases (41.5%), followed by deaths in public places with 332 cases (41%). Additionally, 113 individuals (14%) died in a hospital, and 29 individuals (3.6%) died in prison. Out of the 16 cases that died due to COVID-19 with a history of hospitalization, 8 cases (56.3%) were discharged from the hospital with personal consent and were cared for at home. These individuals had an average of 9 days of hospitalization in the isolated and general wards of the hospital, and 6 individuals experienced respiratory distress during hospitalization. Among those who died in the hospital, 8 cases (7%) had a positive test, and among those who died at home, 23 cases (6.8%) had a positive test. Meanwhile, 8 individuals (34.8%) among those who died at home and had a positive test also reported a history of hospitalization.
The average interval between hospital discharge and death at home was 15.1 days (ranging from 1 to 35 days). It seems that the delay in timely hospital admission led to patients being in critical condition, increasing the severity of the disease and resulting in higher mortality (
30,
31). A study by Birkmeyer et al. revealed that prompt medical interventions and early hospital presentation are associated with decreased rates of severe adverse outcomes and mortality in COVID-19 (
32). None of the prisoners who died in prison had a positive test. Upon entering prison, all individuals were screened for COVID-19, and positive cases were immediately isolated and treated. Therefore, all deceased cases were identified and treated before death. However, environments with excessive crowding and limited access to healthcare, such as prisons, are prone to outbreaks of infectious diseases, underscoring the importance of heightened surveillance and preventive measures (
33) Reports presented worldwide indicate that individuals with a history of COVID-19 can also become re-infected with the virus if exposed to cases of COVID-19 (
29,
30). In this study, 12 cases (70.6%) of the 17 deceased individuals who had a history of contracting COVID-19 and were referred to forensic medicine after death tested positive. Meanwhile, out of 769 cases without a history of COVID-19, 24 cases (1.3%) tested positive after death (P < 0.001). This finding was consistent with the results of Singh et al., which demonstrated that viral mutations and variations in individual immune responses contribute to differences in reinfection rates and that a single infection does not confer complete immunity (
34).
It seems that some individuals develop a false sense of security after contracting the disease and subsequently expose themselves to high-risk environments with high viral loads without observing protective measures (
35). Conversely, the virus's genetic diversity and high mutation rate, particularly in individuals with immune system defects, create conditions that enable the virus to evade immune responses (
36,
37). Among patients with SARI, 13 cases (81.3%) tested positive, while only 2.9% of those without a history of SARI tested positive (P < 0.001). According to epidemiological findings collected from 190 COVID-19 patients at the Policlinico Umberto I Hospital in Rome, 63.6% of patients reported an influenza-like illness (ILI) in their clinical history 1 to 3 weeks before the onset of symptoms associated with COVID-19. The ILI can be considered a risk factor for contracting SARS-CoV-2 in the future (
38). According to the statements of the relatives of the deceased, out of 810 deceased cases, only 4 cases (0.49%) had a history of contact with a COVID-19 patient, of which 2 cases (50%) tested positive. It should be noted that, since the relatives of the deceased were asked about the deceased person's contact history, some relatives may not have been aware of the actual contact history. In the study by Chen et al., 44 individuals (12%) of the deceased had a history of close contact with people confirmed to be infected with COVID-19 (
31).
In the study by Cheng et al., 5.5% of family contacts, 2.8% of non-family contacts, and 25.3% of individuals exposed through healthcare systems were infected with COVID-19 (
39). In our study, the average time interval between death and sampling was 32.6 hours for those who tested positive and 31.9 hours for those who tested negative. No significant difference was observed in the time interval between death and sampling between the two groups (positive and negative test deaths) (P = 0.942). Factors such as viral load at the time of death and sampling conditions appear to be more decisive determinants in the test results.
In our study, the RDT result of 7 individuals (0.86%) among the deceased who died more than 24 hours earlier was positive, and among them, after more than a week of death, the test result of 2 individuals (0.25%) was reported positive. One case tested positive after 10 days, and the second case after 8.5 days postmortem. The results showed that after more than 7 days, the remains of virus particles could still be detected in corpses. However, there was no significant relationship between the time interval from death to testing and the result (P = 0.153). The identification of viral RNA/antigens in cadavers shows that SARS-CoV-2 RNA can be detected in various tissues of cadavers for an extended period after death, depending on environmental conditions. This may indicate the coronavirus's high resistance to unfavorable environmental conditions (
40,
41). The persistence and infectivity of SARS-CoV-2 in corpses underscore the critical need for meticulous personal protective equipment (PPE) protocols among medical personnel (
42). However, the detection of SARS-CoV-2 RNA within cadaveric tissues does not inherently equate to a risk of disease transmission from the deceased (
43).
In the study by Zhou et al., the longest duration of virus shedding was reported as 37 days in recovered patients and until death in deceased individuals (
44). Additionally, some studies have reported that the duration of virus shedding in patients ranges between 3 and 60 days (
45). There is no evidence that the virus is transmitted through the movement of corpses; however, considering that in the current study traces of the virus were detected in corpses up to 10 days after death, it is possible for transmission to occur during contact with the corpse and exposure. Contaminated body fluids should be minimized by wearing PPE. It is necessary to use appropriate eye protection and masks during autopsies, particularly when procedures that generate aerosols are performed (
46). Invasive autopsies, which require specialized personnel and advanced equipment, cannot be performed in areas with limited resources; therefore, in recent years, less invasive autopsies have been recommended (
47,
48). The average interval between the last dose of the COVID-19 vaccine and the time of death was 66 days among cases with a positive outcome. In these cases, 1 individual (2.8%) had received a single dose, and 3 individuals (8.3%) had received two doses of the vaccine. Thirty-two individuals (88.9%) had no history of COVID-19 vaccination. None of the positive cases had a complete vaccination history (three doses). Although only 4 (2.6%) vaccinated individuals had a positive test result compared to 32 (5.5%) of the unvaccinated individuals, this difference was not statistically significant (P = 0.146).
In the study by Juthani et al., 172 (18%) of 969 hospitalized patients with COVID-19 had received at least one dose of the COVID-19 vaccine during hospitalization. Of these, 103 individuals received one dose, 15 received two doses, and 54 received three doses of the vaccine (complete vaccination) 14 days before the PCR test result was positive. Vaccination against COVID-19 effectively prevents infection with COVID-19 or hospitalization (
49). Vaccination is increasingly considered the ideal intervention against infectious diseases (
50). Coronavirus disease 2019 vaccines have played a crucial role in controlling the COVID-19 pandemic, particularly in reducing the incidence of severe cases and deaths caused by the disease (
51). The availability of vaccines is a crucial factor in reducing the number of new infections. Factors such as low education level, female sex, young age, and low income levels can reduce individuals' willingness to get vaccinated (
52,
53). In the present study, 48.9% of the participants were illiterate, 46.7% had a primary to high school education, and only 3.3% had a university education. Additionally, in this study, hypertension and cardiovascular diseases were the most frequent underlying conditions, affecting 90 individuals (11.1%), followed by diabetes in 12 individuals (1.5%) and cancer in 8 individuals (1%).
Thirteen individuals (1.6%) had other underlying diseases; among the 810 deaths referred to forensic medicine, 123 cases (15.2%) had at least one underlying disease. No significant relationship was found between the presence of an underlying disease and a positive test result for COVID-19 in deceased cases (P = 0.83). In two separate studies by Zhou et al. and Wichmann et al., cardiovascular diseases and high blood pressure were reported as the most common underlying diseases among the deceased (
44,
54). Having a chronic disease increases the likelihood of death due to COVID-19 (
28). In the historical cohort study conducted by Zhou et al., all patients over 18 years of age who were hospitalized with laboratory-confirmed COVID-19 at Jinyintan Hospital and Wuhan Lung Hospital (Wuhan, China) and had been discharged or died by January 31, 2020, had an average age of 56 years (range: 18 - 87 years). Of the 191 patients studied, 137 were discharged, and 54 died. Among these cases, 91 patients (48%) had at least one underlying disease. The most common underlying diseases were high blood pressure in 58 individuals (30%), diabetes in 36 individuals (19%), and coronary artery disease in 15 individuals (8%) (
44).
In our study, information related to the underlying diseases of the deceased cases was collected from their relatives. Considering that the information provided by relatives about the deceased may have been incomplete, as well as the lack of access to the relatives of several deceased individuals — such as non-Iranians and homeless people — and the lack of response due to the psychological conditions of those around them, it seems that the prevalence of underlying diseases is lower than in the general population. According to the data from the national survey of risk factors of non-communicable diseases (STEPS) report of 2021 in Sistan and Baluchistan province, the prevalence of high blood pressure was 27.8% (24.9 - 30.7%), cardiovascular diseases 4% (2.8 - 5.2%), and diabetes 13.8% (10.6 - 17%), which differ from the prevalence of these diseases among the deaths referred to forensic medicine. Furthermore, this study is predicated on forensic medical data, which may not fully represent the broader population affected by COVID-19. The study by Tchicaya et al. demonstrated that geographical, social, and cultural disparities, coupled with variations in access to healthcare, result in differential levels of vulnerability across diverse communities (
55). The inadequacies in COVID-19 mortality registration systems have led to incomplete documentation of causes of death during the pandemic. Factors such as insufficient testing, inaccurate recording, and the indirect impacts of socioeconomic determinants contribute to underreporting COVID-19 statistics and limit the generalizability of findings (
56). Similarly, in the present study, the inaccessibility of data regarding specific demographics, including non-native populations, homeless individuals, and the effects of geographical and cultural variations, could potentially influence the study's findings.
Chen et al. studied 799 patients, including 274 confirmed moderate-to-severe or critically ill patients with COVID-19 who were admitted to Tongji Hospital. One hundred and thirteen of these patients died due to COVID-19, and 161 patients fully recovered and were discharged. The average age of the patients who died was 68 years, which was significantly older than that of the patients who recovered. The male gender was more prevalent among deceased patients (83 individuals; 73%) than among recovered patients (88 individuals; 55%). Overall, 71 (63%) of patients who died and 62 (39%) of those who recovered had at least one chronic disease. High blood pressure, cardiovascular disease, and cerebrovascular disease among the patients who died were observed in 54 (48%), 16 (14%), and 4 (4%) individuals, respectively, whereas among the recovered patients, these conditions were observed in 39 (24%), 7 (4%), and 0 (0%) individuals, respectively. The average time from the onset of symptoms to death in deceased patients was 16 days, and the average hospitalization time to death was 5 days. Chronic high blood pressure and other cardiovascular diseases were more common among deceased patients than among recovered patients (
31). In the study by Huang et al., which was conducted on patients with laboratory-confirmed 2019-nCoV infection with positive RT-PCR, less than half of the patients had underlying diseases (32%), which were diabetes (20%), high blood pressure (15%), and cardiovascular diseases (15%), respectively (
57). In the study by Wichmann et al., 12 cases of death with COVID-19 were autopsied between one and five days after death; the average age of the deceased was 73 years. All samples had underlying diseases. Coronary heart disease and asthma or chronic obstructive pulmonary disease (COPD) were the most common comorbidities (50% and 25%, respectively). Additionally, all deaths occurred in the hospital (
54). The mortality rate of COPD patients with a severe form of COVID-19 (hospitalization) ranges from 1% to 62%. In COVID-19, for patients with respiratory diseases (asthma, chronic respiratory diseases, COPD, acute respiratory distress), the probability of death and adverse outcomes is higher. The likelihood of death among hospitalized patients with COVID-19 with an underlying disease was twice as high as that of individuals without an underlying disease (
48).
5.1. Conclusions
Developing community awareness about the importance of timely and early referral to the special care department for high-risk populations is crucial. Due to the persistence of virus particles for more than a week in the deceased and the positive test results observed in this study, it is suggested that personnel working in forensic medicine and all individuals who deal with the deceased in any way during epidemics of infectious diseases use less invasive methods and employ appropriate PPE during autopsies and any actions performed on corpses.
Less invasive methods are safer for health personnel, especially in infectious disease settings, and can be easily performed by trained technicians. Performing a non-invasive autopsy (including blood and cerebrospinal fluid sampling and collecting tissue samples from organs using biopsy needles, followed by histological and microbiological analyses) is a reliable tool for determining the causes of death. Especially in areas where the occurrence of infectious diseases is high, it can be used for prioritization and health planning for the vulnerable populations of countries.