There was one other way forward. “Hans Petrie kept talking about a missing heart,” recalls Cassiman. Petrie knew that during the autopsy on the boy who died in the Temple in June 1795, his heart had been stolen. DNA analysis on this heart might show whether the orphan of the Temple was maternally related to Marie-Antoinette and, if confirmed, put an end to the theory of a substitute. Cassiman was tantalized by the fact that he now had the mitochondrial DNA signature of the Habsburg maternal line—it would be so easy to compare. But where was the heart?
By chance, shortly after announcing his results on Naundorff in 1998, Cassiman received a phone call from an eminent French historian in Paris called Philippe Delorme. Delorme introduced himself, and it was soon clear that he had a deep interest in the case and had studied the life of Louis XVII closely. He knew exactly where the heart could be found and was sure he could get a piece. And he was right.
With painstaking research, Philippe Delorme had traced the whole incredible story of the child’s heart: Pellatan’s “pious theft” during the French Revolution, his Herculean efforts to return the heart to the royal family, Tillos’s theft from Pellatan, the dramatic loss of the heart in the July Revolution of 1830, and its eventual transfer to the Château de Frohsdorf, where it was again caught up in war with the advances of first Hitler’s troops and then the Soviet army. However, during World War II, Delorme told Cassiman, the heart had been rescued by the duke of Madrid’s descendents and ended up in the care of his granddaughter, Princess Marie-des-Neiges Massimo. Eventually, in April 1975, the princess had returned the boy’s heart to France. In a solemn ceremony, the heart had been given to the Duc de Bauffremont, who runs the Memorial at Saint-Denis, the private
organization that oversees the royal graves. More than 150 years since Pellatan had first tried to place the relic in the royal crypt, the heart of the orphan of the Temple had finally arrived at the Basilica at Saint-Denis. It had been placed in an underground chapel in the crypt,
La Chapelle des Princes,
and, said Delorme, it is still there to this day.
After speaking to Cassiman, Delorme immediately went to see the Duc de Bauffremont. He explained that his historical research had shown that it was indeed the same child’s heart from the Temple prison in 1795 as the one now in the Basilica. Despite its astonishing past, this stolen heart, the only vestiges of the child who died in the Tower, could yet reveal the true story of the dauphin. The Duc de Bauffremont was intrigued and agreed to the tests.
In June 1999, the three men gathered round the tiny heart in its urn hidden away in a side chapel at Saint-Denis. For Philippe Delorme, it was an emotional moment as the wrought-iron gate to
La Chapelle des Princes
was unlocked and they walked carefully in the half-light between the dusty coffins, seeming almost to walk through time itself as they travelled back two hundred years, reaching the shelf on the far wall where the crystal urn with its secret waited. “It was scarcely like a heart, it was so small and dry, and yet, at the same time, it was such a powerful symbol,” thought Delorme. True scientist, Cassiman remained unmoved at the sight—to him it was nothing more than a biological specimen. The urn was sealed, so he could not examine the heart directly; he suspected that it was in bad shape, given its eventful history. Delorme asked what their chances were of obtaining any DNA. Cassiman could guarantee nothing, but agreed they would at least try.
There were earnest discussions about the protocol, permissions and the nature of the genetic tests; because of the public interest in the case, they decided notaries should record every detail of the procedure. Cassiman agreed to look for another genetics team that could also carry out independent tests. “They asked me specifically—don’t take a Frenchman.” Cassiman smiled. “There were too many vested interests in the story in France!” He nominated a German laboratory at Munster University, led by Professor
Bernard Brinkmann. Brinkmann’s team was one of the few in Europe that had considerable expertise in the use of mitochondrial DNA in forensic cases.
With arrangements complete, on December 15, 1999, after a short ceremony at Saint-Denis, the boy’s heart in the crystal urn was placed in a hearse and travelled through the streets of Paris once more on its final journey. At the nearby Thierry Cote Medical Laboratory, Cassiman soon found his initial doubts were not justified. It was possible to see every detail of the heart, the muscular structure, the coronary arteries, all the vessels and compartments; it was extremely well preserved. However, time had turned it to stone and it could only be cut with a saw. Doctor Els Jehaes severed the bottom tip of the heart and also took a sample of the aorta, the great artery coming from the heart. These segments were then carefully divided: one specimen for Cassiman and his team in Belgium, the other for Professor Brinkmann in Germany, and then the jars were sealed.
Cassiman himself carried the five hundred milligrams of the boy’s heart back to Leuven, past the old town and Maria-Theresiastraat to the Center for Human Genetics on the edge of town. He took the sealed tube up to the sixth floor, along gleaming corridors lined with canisters of gas, fridges and freezers. It was here that the carefully labelled locks of hair from Marie-Antoinette were stored, along with the locks from the dauphin’s aunts. Now the segments of the child’s heart, too, joined them in a freezer awaiting examination.
Once the notary arrived to witness the breaking of the seal of the jar in Leuven, the heart was taken to the extraction laboratory. To get at the child’s DNA, first they had to destroy the heart tissue, removing cell walls, proteins, and any other material. The fragments of heart were cut into very small pieces with a sterile saw and then crushed, either with a pestle and mortar, or frozen in liquid nitrogen and broken up with a hammer. “It was like grinding meat,” says Cassiman. Gradually, the child’s heart was reduced to a fine dust in a sterile tube. Standard chemical procedures were used to clean it and break down the cell walls with special enzymes. To Cassiman’s surprise, the heart contained relatively high amounts of DNA. As before, this
was put in the PCR machine to copy the genetic signature of the child millions of times. These samples were then placed in the sequencing machine, to read the order of the 1,100 bases in the two key sections of mitochondrial DNA.
In the sterile modern laboratory, under the bright fluorescent lights, faint echoes from another age emerged as the secrets of the heart finally yielded to modern science. Cassiman was able to obtain three identical sequences of mitochondrial DNA from the child’s heart—the essential clue to his identity.
“As soon as we had the little boy—we could see his sequence was identical to the living relatives,” says Cassiman. To double-check, they compared it manually—base by base. It was an exact match. “We had worked for years on the Naundorff case and come out with a result which I was not one hundred percent pleased with scientifically,” says Cassiman. “Our results had made the point that Naundorff was not Louis XVII—but not to our satisfaction. We had the problem of the two additional changes in the mtDNA in the living relatives of Marie-Antoinette, compared to the sequences from Marie-Antoinette’s and her sisters’ hair. Now we had the heart, everything fell into place because it matched the living Habsburg relatives perfectly! This confirmed for me that we had not got a complete result from the older relatives.”
There was just one final test. Could he get the Habsburg sequence found in the boy’s heart and Marie-Antoinette’s living relatives from the older biological specimens—the hairs of Marie-Antoinette or her sisters? They had spent some time developing their methodology and developed a probe that would allow them to amplify shorter sequences in the PCR machine. The original tests had been done on longer DNA fragments that were over 250 base pairs. They now recognized that it was easier to accidentally include a contaminant in the longer fragments, and miss some of the key positions that make up the true sequence. With the new probes, they could break the DNA into tiny fragments, a mere 100—200 base pairs, to obtain a more accurate result.
The hairs of Marie-Antoinette herself did not contain sufficient DNA to
get a full result. But in the freezer outside the laboratory they had just one hair shaft left—from her sister Johanna-Gabriela. “I felt if we could confirm the same sequence that we found in the living relatives in that one hair then I would be satisfied,” declared Cassiman.
He repeated the cycle: extraction, analysis, PCR and sequencing. Once he had the results he sent them to the computer in his office where he had stored the results from the boy’s heart. Here in his orderly office, sunlight streaming through the windows, classical music playing softly as he worked, Cassiman could study the result, base by base. The distinctive genetic signature from the single hair of Marie-Antoinette’s sister—the Habsburg maternal line—was lined up against the heart of the son she had lost. Using the ghostly imprint of the genetic material of mother and son, united for the first time in two hundred years as an electronic signal in the laboratory computer, together they were able to reveal what really happened. The sequence from Johanna-Gabriela, the aunt the dauphin had never met, was an exact match to the heart of the boy. The DNA signatures for the critical region of mitochondrial DNA were
identical.
On April 3, Professor Brinkmann came to Leuven to compare data. “They had the same results—and that was interesting,” Cassiman said, with classic understatement. “Not only did we have sequence alignment and identity from the heart with living and deceased relatives, but we also had independent confirmation from another lab.” From a scientific point of view, the puzzle was solved. Meanwhile, Delorme had been waiting anxiously in Paris. He was delighted when a fax finally arrived from Cassiman confirming all of his historical research.
On April 19, 2000, Professors Cassiman and Brinkmann, Philippe Delorme and the Duc de Bauffremont assembled in Paris to hold a press conference to announce their findings. Members of both lines of the Bourbon family were present; Prince Louis de Bourbon, duke of Anjou, one of the closest living relatives of Louis-Charles, arrived from Spain for the occasion. There were also representatives of Naundorff’s family. A large crowd of TV crews, international press, and photographers had gathered to hear the results.
Cassiman and Brinkmann carefully outlined the steps in the scientific testing. “The comparison of the DNA appeared to show beyond all reasonable doubt that the heart came from a child that was maternally related to the Habsburg family,” Cassiman told the reporters. “The sequence for the Habsburg family is unique, since it has not been observed in a collection of more than one thousand seven hundred European mtDNA sequences. All this, taken with the historical record, provides strong evidence to support the proposition that this is the heart of the lost dauphin.”
With his customary scientific caution, he also explained the limitations of the testing. “The science does not
prove
beyond doubt that the heart belonged to Louis XVII, or even to a son of Louis XVI and Marie Antoinette,” he said. “The scientific tests can only show that the heart in the crypt has to belong to the son of a maternal relative of the Habsburg family.” When asked whether it was possible that the heart could have come from some other unknown son of Marie-Antoinette herself, he replied, “Marie-Antoinette may have had a relationship with the Swedish count—Axel Fersen—but there is absolutely no historical evidence of a secret son who died at the same age, whose heart could have been taken in this way. Scientifically, I can’t disprove this, but it does not fit with the historical data.” Marie-Antoinette’s life was so public, another child would have been impossible to conceal.
There was also the question of whether the heart could have belonged to the older son of Marie-Antoinette, the first dauphin, Louis-Joseph, who died of illness in Versailles before the revolution in 1789 at the age of eight. “This too was unlikely,” Delorme reasoned, “because the heart removed from Louis-Joseph was embalmed. When a heart is embalmed it is cut open down the middle and filled with herbs and embalming liquid.” Yet the heart on which Cassiman conducted his tests had clearly not been embalmed; the aorta was cut roughly and the tissues were fragmented and dried as is consistent with the heart’s tumultuous history. Consequently, he explained, although Louis-Joseph’s heart was lost after the Revolution and has never been found, it is most unlikely to be from Marie-Antoinette’s older son.
Some reporters asked about the independence of the tests, given that they
were paid for by the royal family trust. Cassiman laughed at this suggestion; he was paid around 1,500 pounds, which barely covered laboratory costs, and he had undertaken the research primarily for the scientific challenge. He pointed out there are only a few forensic laboratories in Europe that can test mtDNA; the two laboratories were specifically chosen because they were not French and had no interest in the outcome. Others still speculated that the extraordinary history of the heart must cast doubt on the findings. Professors Cassiman and Brinkmann dismissed this criticism, explaining that two independent teams had come to exactly the same result.
“And is this ninety-nine percent sure?’ pressed the reporters.
“Yes, I’m satisfied,” replied Cassiman. “If you take our results with the historical evidence—and if that is correct—then our results are one hundred percent. Everything seems to indicate that it was the young dauphin who died alone in the Temple prison in tragic circumstances.”
“This is the end of two hundred years of uncertainty,” declared Delorme. “It puts to an end a mystery that has absorbed so many of us. The DNA analysis shows the child’s heart is from a member of the Habsburg family. The historical research shows that this heart came from the orphan of the Temple. Since, apart from Marie-Thérèse who survived, the only other relative of Marie-Antoinette in the Temple in 1795 was Louis-Charles, now we have an answer. It was Louis XVII, the little king of France without a crown, who died in the Temple prison. It’s definitive.”