The first confirmed case in Sao Paulo state was February 26, 2020. International importations accounted for a majority of cases at the outbreak’s start before local cases were detected(25). Following the outbreak’s spread to local community transmission, there were four major policy changes in the state’s RT-PCR testing guidelines. From March 17 to June 30, 2020, state surveillance policies limited RT-PCR use to hospitalized and symptomatic individuals and health professionals. Under Testing Policy I, an average of 30 tests per 100,000 was conducted, and an average positivity rate of 25.67(%) was observed (Table 1). When São Paulo state expanded RT-PCR testing to permit testing of non-hospitalized, symptomatic cases (Policy II), an average of 166.23 tests per 100,000 were performed, and the positivity rate of 32.21(%) was observed from July 1, 2020 to September 14, 2020. There was a significant increase in COVID-19 cases during this period with a peak number of the average cases registered per 100,000 in weeks 32 and 33 of 2020. On September 15, 2020, the Brazilian Ministry of health expanded the notification criteria, including clinical and epidemiological criteria (Policy III). This could have resulted in an expansion of the testing effort for asymptomatic and suspicious cases. However, an average of 197.41 tests per 100, 000 between September 15, 2020 to February 25 2021 were registered. In this period, the average positivity rate of 31.22(%) ranged from a weekly average rate of 19.76(%) (week 45, 2020) to 40.58(%) (week one, 2021) (Figure 1, Panel a). The second highest peak in cases was observed in weeks two and four of 2021. When guidelines were revised to incorporate genomic surveillance as part of the state’s RT-PCR testing strategy from February 26 to June 5, 2021 (Policy IV), an average of 305.52 tests per 100,000 were conducted, and an average positivity rate of 41.80(%) was observed. This period coincides with the third highest peak in cases observed in week 22 of 2021.
Significant variation in the volume of tests conducted across the state’s 17 Regional Health Departments (RHD) over the past fifteen months is observed. From March 1, 2020 to June 5, 2021, a weekly average of 168.41 RT-PCR tests per 100,000 were performed across the state. São José do Rio Preto was the RHD with the highest average of tests performed (322.07 tests per 100,000), while Grande São Paulo was the lowest average of tests conducted (74.18 tests per 100,000). There has also been a substantial variation in the positivity rate across the regions. Throughout the period, the state average positivity rate was 31.21(%). Araraquara presented the lowest average positivity rate (23.16%) and Registro the highest average positivity rate (40.55%).
In the early months of the pandemic, public health testing efforts were coordinated by the Instituto Adolfo Lutz. In April 2020, a new public laboratory system was created with the Instituto Butantan as its central manager (26) to coordinate efforts to complement efforts undertaken in the IAL network. In this period, university laboratories and two private labs were incorporated into the public laboratory network to expand testing. Notwithstanding the expansion of the network and central management, public laboratories remain unequally distributed across the São Paulo state territory. Only 12 of the state’s 17 Regional Health Departments (RHD) have at least one local laboratory in their region. While Grande São Paulo has four local laboratories (Instituto Adolfo Lutz Central, Instituto Adolfo Lutz Santo André, Instituto Biológico, and Instituto Butantan), five RHD do not have a local laboratory (Barretos, Franca, Registro, São João da Boa Vista, and Taubaté). There was considerable delay in expanding public health laboratory capacity in the state. Initially, RT-PCR tests were limited to processing 1,200 tests per day in the Instituto Adolfo Lutz (IAL) lab (Figure 1, Panel b). By September 2020, the public laboratory network had expanded its processing capacity to 13,154 RT-PCR tests per day. In February 2021, the state reached a capacity to process 17,394 tests per day. No further increases were observed after this date.
The state’s public laboratories processing capacity is also unevenly distributed. Some laboratories have still not acquired the capacity to process tests (Instituto Adolfo Lutz Araçatuba and Instituto Adolfo Lutz Campinas). The two public laboratories with the highest daily processing capacity (Instituto Adolfo Lutz Central with a capacity of 1,100 tests and the Instituto Butantan with 4,500 tests) are in the same RHD (Grande São Paulo). The State of São Paulo also contracts private laboratories to process RT-PCR tests. These providers (DASA and Grupo Fleury) were contracted to process 1,945 tests per day in 2020. In 2021, private laboratories were contracted to process 3,545 tests per day (20.1% of daily testing in the network). In contrast, there was a smaller increase in the IAL network testing capacity (2,304 tests per day in September 2020 to 2,895 tests per day by February 2021). Data are only published on the date and the lab that received the RT-PCR test. The lab processing the test and the test result date are not informed in public data sources.
Between March 1, 2020, and June 5, 2021, 1,763 samples were collected for genomic sequencing from RT-PCR positive samples by the Instituto Adolfo Lutz network in the state of São Paulo (18.06% of the 9 761 samples collected and 20.65% of the 6,324 genomes sequenced in this period in GISAID for São Paulo state). However, given delays in uploading sequences to the registry database and missing values, only 1,299 of these genomic sequences were deposited with GISAID in the same period. On average, 0.0024 of RT-PCR positive tests in each RHD in the state of São Paulo were sequenced per week. In 2020, the number of sequences registered in GISAID was limited. On average, 0.70 sequences per RHD were registered per week in 2020 (based on 0.0025 of RT-PCR positive tests). While, on average, 3.32 sequences were registered per week in 2021 (based on 0.0023 of RT-PCR positive tests). The highest weekly average was observed in week six, 2021, when 9.12 genomic sequences were performed across the RHDs. Most of the sequences were uploaded to GISAID with considerable delay. The time lag between the specimen collection and submission has been remarkably high in 2020 (on average, 149.3 days). Since March 2021, there has been a marked rise in sequencing and submission. The time lag has substantially decreased in 2021 (54.4 days from specimen collection to submission).
Genomic sequencing by the IAL varies considerably across the state’s RHDs (Figure 2). Grande São Paulo was the RHD with the highest weekly average of genomic sequencing performed (on average, 9.54 per week), while Franca was the lowest average of genomic sequencing conducted (on average, 0.33 per week). The lag in time from collection to submissions of the SARS-CoV-2 genome to GISAID by the IAL network has also varied significantly across RHDs. In 2020, the lowest average weekly time was 97.50 days from samples in São José do Rio Preto, and the highest average was 210 days from samples collected from Baixada Santista. In 2021, this period was considerably reduced. The IAL network performed the lowest average weekly time in Registro (41 days) and the highest average in Baixada Santista (68 days). The available data do not permit verification of whether the difference in processing time is due to which laboratory of the network is performing sequencing.
For the state overall, we transformed one of these dependent variables (RT-PCR tests Positivity Rate) into first differences of their natural logarithm, which is the per-week growth rate (Table 2). The results showed a 0.005 [95(%) confidence interval (CI) -0.003; 0.013] increase in the log of RT-PCR tests per 100,000, -0.0008 [95% confidence interval (CI) -0.0058; 0.0042] decrease in the ratio of RT-PCR tests per case, and a -0.0003(%) [95(%) confidence interval (CI) -0.0016; 0.001] decrease in the growth rate of the RT-PCR test positivity rate following Testing Policy II (Figure 3, Panel a). In turn, Testing Policy III, results showed a 0.005 [95(%) confidence interval (CI) 0.002; 0.007] increase in the log of RT-PCR tests per 100,000, a 0.0076 [95(%) confidence interval (CI) -0.0046; 0.0197] increase in the ratio of RT-PCR tests per case, and an 0.002(%) [95(%) confidence interval (CI) 0.0004; 0.003(%)] increase in the growth rate of the RT-PCR positivity rate. As a result, Testing Policy II did not produce a statistically significant increase in surveillance efforts. Following Testing Policy III, there was an increase in the mean volume of testing in each RHD, but the test positivity rate increased due to insufficient testing expansion.
Uneven improvements in testing outcomes were observed across the state (Figure 3, Panel b). Following Testing Policy II, an increase in the log of RT-PCR tests per 100,000 was only observed in four RHDs (RHD V, VIII, XI, and XVI) and a reduction in the log of RT-PCR tests positivity rate occurred in six RHDs (X, XI, XII, XIII, XIV, and XV). With respect to the ratio of RT-PCR tests per case, a decrease was observed in four RHDs (III, XIV, XV, and XVII) and an increase in five RHDs (I, II, VII, XIII, and XVI) after Testing Policy II. Under Testing Policy III, there was an increase in the log of RT-PCR tests per 100,000 in eight RHDs (I, III, VI, IX, XI, XII, XIV, XVI), and no reductions in test positivity rates were observed. Instead, test positivity rates increased after Testing Policy III in two RHDs (IX, XII). After Testing Policy III, the ratio of RT-PCR tests per case increased in two RHDs (VIII and XIII). These results are only with respect to those RHDs in which no evidence of first or second-order autocorrelation was found in the residuals after using Newey-Standard Errors for first-order serial correlation.
The impact of genomic surveillance efforts was analyzed based on the number of samples and the time delay between sample collection and deposit to the GISAID (Table 3). Testing Policy IV did not produce a statistically significant increase in genomic surveillance efforts. Results showed a -0.020 [95(%) confidence interval (CI) -0.046; 0.005] decrease in the number of samples and a 0.028 [95(%) confidence interval (CI) -0.142; 0.198] increase in the time delay between sample collection and deposit to the GISAID platform after the introduction of the policy.
Considering the results related to the four analyzed policies, there is no significant association between the public RT-PCR testing variables and each policy. Figure 1 also shows that there was no improvement in the testing effort in the period studied, even though the previous policies were maintained and could have shown accumulated positive effects, with an improvement in all RT-PCR testing effort variables.
RT-PCR testing intensity and outcomes are correlated with testing policy interventions, population density, income per capita, the proportion of the population dependent on SUS and social vulnerability. RT-PCR tests per 100,000 are negatively associated with high socioeconomic vulnerability (p-value=0.000) and with a higher population density (p-value=0.000). RT-PCR test positivity rate is positively correlated with high socioeconomic vulnerability (p-value=0.000) and with a higher population density (p-value=0.013), and it is negatively correlated with average income per capita (p-value=0.026) (see Figure 4). COVID-19 cases, in turn, are negatively correlated with high socioeconomic vulnerability (p-value=0.027) and higher population density (p-value=0.001). The number of genomic sequences collected and uploaded at the GISAID is positively correlated with high socioeconomic vulnerability (p-value=0.025), higher population density (p-value=0.000), and income per capita (p-value=0.000), and it is negatively associated with a higher population dependent on SUS (p-value=0.000). The average time delay between sample collection and deposit to the GISAID is positively correlated with high socioeconomic vulnerability (p-value=0.014), population density (p-value=0.000), and average income per capita (p-value=0.002).